Plant like starches and the method of making them in hosts

ABSTRACT

This invention relates to hosts containing constructs with genes from the starch pathway. More typically the present invention relates to bacterial hosts that form plant like starches. Additionally the present invention relates to plant hosts that have genes from the starch pathway. The invention further relates to the starches produced by said hosts.

[0001] The present application is based on U.S. Provisional Application No. 60/062,939, filed Apr. 4, 1997, the entire contents of which is incorporated hereby by reference.

BACKGROUND

[0002] 1. Field of Invention

[0003] This invention relates to hosts containing constructs with genes from the starch pathway. More specifically the present invention relates to bacterial hosts that form plant like starches. Additionally the present invention relates to plant hosts that have genes from the starch pathway. The invention further relates to the starches produced by said hosts.

[0004] 2. Description of Prior Art

[0005] The starch using industry includes diverse industries such as candy makers, makers of adhesives and paints, gravy makers, paper producers, etc. Since the demand for starch, (which is formed of amylose and amylopectin), has been dramatically increasing for specialized food and industrial uses, efforts have been undertaken to tailor the quantity and quality of starch for specific food and industrial uses.

[0006] This industry has overtime looked for a number of different starches having, high viscosity, lower viscosity, higher gelling strength and lower gelling strength different boiling points etc. Each starch tailored for a number of uses. The industry has utilized mutant starches that have less amylopectin and mutant starches with more amylose for tailored specifications. For example the increased amylose starch has been used in the gelled candy making area. And the industry has used the increased amylopectin starches formed by mutants such as wx and wx su2 containing little amylose and mostly amylopectin for thicken foods like pudding, pies, gravies, frozen foods and batters, stews, canned foods and baby food. Additionally the mutant starches of different types have usefulness as adhesives and as sizing.

[0007] The other method used to address the industry needs for tailored starch is the use of chemical modification of the starch. Chemical derivation of the starch are produced by chemically reacting the starch with the monofunctional reagents to introduce the substituents such as phosphate, acetate, succinate groups to stabilize the starch. Unfortunately, these types of starches can be subjected to government regulation and additionally have less acceptance generally due to the added cost of the starch.

[0008] Starch is the major form in which carbohydrates are stored in biological systems. Plant starch in chloroplasts is transitory and storage starch accumulates in storage organs of many plant. Starch can be found in all organs of most higher plants including leaves stems and roots and fruits and embryo and endosperm. In addition to higher plants starch, similar polysaccharide (glycogen) has been found in bacteria. Many bacteria produce a reserve polysaccharide similar to the glycogen found in animals.

[0009] Storage polysaccharide has been classified as being in two groups, group one has storage in the cytsol of the cell and the second group within the plastid. Escherichia coli produces a polysaccharide within the cytsol. Starch producing plants typically store starch in the plastids. Typical starch bearing plants include cassaya, potato, corn, peas, rice, wheat, barley. The main starch storing tissue of corn, rice wheat and barley and oats, the cereal grains, is the endosperm.

[0010] Starches are also classified by the plant source, for example cereal starches are from cereal grains such as maize, rice, wheat, barley, oats and sorghum; tuber and root starch are from potatoes and yams and cassaya.

[0011] The pathway of starch synthesis is not well understood. Generally, as noted above starch from plants, consists of two major components: amylose and amylopectin. These intertwine in the starch granule of the plants. Amylose is a linear polymer of alpha 1-4 bonded anhydroglucose units while amylopectin is a branched polymer comprised of linear chains alpha 1-4 linked anhydroglucose units with branches resulting from alpha1-6 linkages between the linear chains. It has been known for sometime that mutant genes in starch bearing plants can be expressed and that the properties of the starch can be altered. The proportion of the two components and their structures in the mutant primarily determine the physical-chemical properties of the starch.

[0012] Thus the lack of a clear understanding of the starch synthesis pathway and the difficulty of employing mutants limited the industry to the use of existing and producible mutant starches (cereals containing mutant starch can show a tremendous yield penalty in field environments) or to the chemical modification that could be made to the starch. In the last decade the industry has been studying the effects of certain starch genes in plants and bacteria in an attempt to more clearly understand starch synthesis. Since the late 80's it has been possible to transform plants and bacteria to contain isolated genes. In response to this the industry has transformed potatoes with a bacterial gene GS and with starch soluble synthase III gene in the antisense (forming a mutant). As part of these potato starch experiments bacteria has been transformed with certain potato starch genes. For example the SSSIII gene from potato was transformed into E. coli deficient in glgA gene. The effect of glgC and branching enzyme I and II in combination in a mutant E. coli has also been studied and glycogen like product was reported. The starch industry that is commercial does not have a particular interest in the production of glycogen which is the polysaccharide produced by bacteria and animals (the health care industry may have some such interest). The industry has thus not yet been able to generate tailored starches at reasonable prices through plant gene transformation. There remains a need for the industry to find new starches that are useful due to their changed characteristics such as lower viscosity and new starches that are useful because of their higher viscosity and new methods of producing such starches.

[0013] Glycogen synthesis in E. coli and starch synthesis in higher plants have similar pathways involving ADPGlc pyrophosphorylase, starch synthase,(SS) or glycogen synthase (GS,) and branching enzyme (BE). It has been suggested that ADPGlc pyrophosphorylase plays a pivotal role in regulating the amount of starch synthesized, while starch synthase and starch branching enzyme primarily determine the starch structure. Multiple forms of SBE and SS have been identified in many plants including maize, rice, pea and potato. In addition to the waxy gene coding for granule bound starch synthase (GBSS), three genes coding for the other forms of SS have been isolated from maize endosperm. Maize is the only cereal crop from which the genes coding for the five forms of SS have been isolated. Clearly a number of these sequences are published and known to those of skill in the art. Genes coding for maize SBE have also been cloned and characterized. Previous reports have demonstrated that maize SBEI has a higher rate of branching amylose than SBEII and preferentially transfers longer chains, while SBEII shows a higher rate of branching amylopectin and preferentially transfers shorter chains. In comparison with SBE, less is known about the specificities and functions of multiple forms of SS. In Waxy maize, which lacks GBSS, only amylopectin is synthesized and amylose is missing. Therefore, it is generally accepted that GBSS, encoded by waxy gene, is primarily responsible for the synthesis of amylose. Study of way mutation in Chlamedomonas reinhardtii has suggested that GBSS is also involved in amylopectin synthesis. Although it has been reported that Chlamedomonas reinhardtii SSII controls the synthesis of intermediate size glucans of amylopectin in Chlamedomonas, direct evidence for the functions of SS in higher plants is still missing. Antisense technology has been used to study the functions of SS in potato, however, the results are inconclusive.

[0014] In an article written by Hanping Guan et al., entitled AMaize Branching Enzyme Catalyzes Synthesis of Glycogen-like Polysaccharide in gig B-deficient Escherichia coli @. Published in Proc. Natl. Acad. Sci. USA, Vol. 92, pp. 964-967, February 1995 Plant Biology a specific glycogen like polysaccharide from a transformed E coli was reported. This article taught the transformation of an E coli bacteria with maize BEI and BEII. These enzymes were transformed into two E coli hosts. One of the bacterial hosts was a wild type and the other was a mutant. The mutation to the bacteria was the reduction of the activity of glycogen BE in the AC71(glgB−) so that the mutant was essentially free of BE activity. The paper analyzed the debranched alpha-glucan isolated from the four different transformants. The first host was E. coli containing glgB and the second host was the AC71 without any transformed genes then AC71 transformed with maize BEII, and then AC71 with maize BEI, then AC71 with maize BEI and BEII. The resultant polysaccharide products were analyzed by HPLC, by chain length and relative peak area and by mole distribution of chains. The study led to the understanding that BEII could play a role in synthesis of the short chains of amylopectin and BEI could be involved with the longer chains of amylopectin. The paper also noted that the mutant host AC71 produced more chains with chain length of 6 then did the wild type E. coli. The paper also noted that the maize BE and the GS of the host did not produce amylopectin like polysaccharides. The article suggested that the concerted action of GS with different BE=s could play an important role in determining the final structure of the polysaccharide synthesized. The article by Guan ends by suggesting that his study had established the basis for studying the concerted actions of BE and SS in a bacterial model system.

[0015] The expression of E coli GS (glycogen synthases) in potatoes showed a large incidence of highly branched starch. This result was published in an article in Plant Physiol. 104,1159-1166 by Shewaker et al. This potato does not appear to be of much commercial use at this time.

[0016] The industry still needs the option of producing plant like starches in a fermentation process from bacteria and thus without the necessity of breeding and growing environment sensitive plants; and the option of producing plants that generate the specific tailored starch through a plant host. And the industry needs altered and new starches that are cereal like starches or root and tuber like starches in large quantities and inexpensively thus avoiding having to use chemical modification of starch. The industry needs a host that can be readily transformed to supply different starches tailored to the industry=s need. Specifically the industry needs a host that supplies various different starches including those not capable of being made in plants or bacteria presently.

OBJECTS AND ADVANTAGES

[0017] Accordingly, several objects and advantages of the invention are to produce plant like starch through the process of fermentation.

[0018] Additional objects and advantages are the production of new starches in plants.

[0019] Still further objects and advantages will become apparent from a consideration of the ensuing description and accompanying drawings.

[0020] Another object of the present invention is the synthesis of polysaccharides including amylose, amylopectin in E. coli, and/or fungal and yeast by plant starch synthesizing enzymes including SS, SBE, bacterial branching enzyme, glycogen synthase and other enzymes in other living organism.

[0021] Yet another object of my invention is using each or combination of these enzymes or modified enzymes studied in this patent to produce or to improve polysaccharides in any living organism including starch synthesis in plants.

SUMMARY OF THE INVENTION

[0022] The invention provides DNA constructs in a host that include most of the genes in the starch pathway of a plant such that the host produces a plant like polysaccharide. And in one embodiment produces maize starch including slightly different embodiments that make specific maize mutant like starch in a non plant host. This invention encompasses a bacterial host containing a combination of two or more of such genes SSI, SSSIIa, SSIIb, SSSIII, GBSS, BEI and BEII when the combination does not form glycogen like material. This invention encompasses a plant host transformed with any of the following maize genes or a plant host having a combination of two or more of the following maize genes SSI, SSIIa. SSIIb, SSSIII, GBSS, BEI and BEII in a hybrid or an inbred rice plant.

[0023] Additionally the present invention includes new polysaccharide produced by a transformed host. The host having a wildtype, which does not produce the new polysaccharide, the transformed host expressing at least two exogenous starch synthesis genes, the genes are selected from a group consisting of starch synthesis genes such as SSI SSIIa, SSIIb, SSIII, GBSS and optionally including at least one of the BEI and BEII genes wherein the transformed host is capable of producing such new polysaccharide.

[0024] The invention also covers a new polysaccharide wherein the host also expresses the exogenous genes selected from the following group consisting of bacterial glycogen inducing genes are selected from the group glgA, glgB, glgC and any mutants thereof. Or wherein the host also expresses the exogenous genes selected from the following group consisting of plant granule bound enzymes. And the new polysaccharide wherein the starch synthesis genes are selected from the group consisting of BEI and BEII.

[0025] The present invention broadly encompasses a host containing a transformed Glg C gene and at least one of the starch branching enzymes genes in a host in combination with at least one other transformed starch gene wherein the host produces a polysaccharide product. And a host containing transformed bacterial gene and at least one of the non starch branching enzymes selected from the group consisting of debranching enzymes and soluble starch synthase

[0026] A method of producing polysaccharides which are non glycogen like in a host comprising transforming a host capable of being used in a fermentation process, with genes selected from the group which produce starch synthesizing enzymes, or glycogen synthesizing enzymes such that the host produces nonglycogen like starch, and employing the host in a fermentation process that produces polysaccharides. The host is bacteria, or a fungal or a yeast. Additionally the method of this invention includes the use of bacterial genes also such as the glycogen synthesizing genes including the glgC, glgA, glgB genes. A method wherein the genes which produce starch synthesizing enzymes include genes encoding for starch soluble synthases I, Ia, IIb and SS III (dull). A method wherein the genes which produce starch synthesizing enzymes include genes encoding for starch debranching enzyme and branching enzymes. Tire invention covers the modified starch synthesizing enzymes including the N-terminally truncated SS.

[0027] In other word the invention covers a host transformed to carry a gene active in glycogen production, and at least one nonstarch branching gene active in the production of at least one of the following polysaccharides amylopectin and amylose in its original host. The host can be a monocot or a dicot plant. The host can be a cereal bearing plant. Or the host can be a bacteria.

[0028] More specifically the invention includes a host wherein at least one nonstarch genes active in the production of at least one of the following polysaccharides, amylopectin and amylose in its original plant, is selected from the group consisting of starch soluble starch synthase I, IIa, IIb, III genes and debranching enzyme gene (su1), GBSS gene, sh2 gene and bt2 gene. A host including at least one of the starch branching enzyme genes such as BEI gene, BEII gene.

[0029] The present invention can also be described as a host transformed to carry a gene active in ADPG production, and at least one starch gene active in the production of at least one of the following polysaccharides amylopectin and amylose in its original host wherein the host produces polysaccharides that are plant like starch and not glycogen like.

[0030] Additionally the host can be transformed to carry a pyrophosphorylase gene, and glycogen synthase gene.

[0031] The scope of the present invention includes a host deficient in alpha 1,4 glucan synthesizing ability and alpha 1,4-1,6 branching enzyme capability transformed to express at least one a plant starch soluble synthesis gene. And the host can also include being transformed to express at least one gene encoding for debranching enzyme, and/or a gene encoding for starch soluble synthase I, starch soluble synthase enzyme Ia, IIb, starch soluble synthase enzyme III. This host can including being transformed to express at least one gene encoding for starch branching enzyme.

[0032] This invention also includes the production of a glycogen like material in plants. Attached hereto are a number of plasmids described by the figures and by table one that are part of the present invention and are claimed herein. One such example is the plasmid wherein the plasmid is in a carrier host and the plasmid contains the SSIIa gene with the n terminus GENVMNVTVV and wherein the gene is approximately 1561 base pairs in length. The invention includes mutant hosts such as mutant plants like waxy rice and potatoes and corn as example and wherein the host is a mutant E. Coli, or fungus.

BRIEF DESCRIPTION OF THE DRAWINGS

[0033]FIG. 1 shows a graph which gives the relative peak area in percent and the chain length of glycogen and starch soluble synthase I (SSI), starch soluble synthase II (SSSIIa), starch soluble synthase IIb (SSIIb). Thus this shows the specificities of Maize SS=s in chain elongation.

[0034]FIG. 2 shows plasmid pEXSC-MBEI with 7661 base pairs and promoter T7 and a Kanamycin gene and glgC and the maize starch branching enzyme I (MBEI).

[0035]FIG. 3 shows plasmid pEXSC3C with 7461 base pairs and promoter T7 and ampicillin gene and the maize starch soluble synthase gene Iia. pEXS3c is the 1082 bp Nde I-EcoRI fragment containing the N-terminus of MSSSIIa (from MSSIIa in pBSK) subcloned into the Nde I-EcoRI sites of pEXS3a, replacing the N-terminus of IIA-2 with the longer Ia N-terminus. MSSIIa is the mature maize SSIIa and is 2090 bp long. The following sites are not contained in the MSSIIc insert: Apa I, BglII, Eco V, Not, Sipe I, and Xba I. The N-terminus of this plasmid is AEAEAGGKD.

[0036]FIG. 4 shows plasmid pEXSC-MBEI-MBEII with 9971 base pairs and promoter T7 and a Kanamycin gene and glgC and the maize starch branching enzyme I (MBEI) and the maize starch branching enzyme II (MBEII).

[0037]FIG. 5 shows plasmid pEXSC-MBEII with 7521 base pairs and promoter T7 and a Kanamycin gene and glgC and the maize starch branching enzyme II (MBEII).

[0038]FIG. 6 shows plasmid pEXSC-3a with 7990 base pairs and promoter T7 and a Kanamycin gene and the glgC gene and the maize N-terminally truncated starch synthase gene IIa (MSSIIa-2). The N-terminal sequence is GENVMNVI.

[0039]FIG. 7 shows plasmid pEXSC-8 with 7079 base pairs and promoter T7 and a Kanamycin gene and the glgC gene and the maize starch soluble synthase gene I and version I-. 2.(MSSI-2), An N-terminally truncated SSI.

[0040]FIG. 8 shows plasmid pEXSC-9 with 7551 base pairs and promoter T7 and a Kanamycin gene and the glgC gene and the maize starch soluble synthase gene IIb (SSIIb). The N-terminal sequence is AAAPAGEE.

[0041]FIG. 9 shows plasmid pEXSC-10 with 7211 base pairs and promoter T7 and a Kanamycin gene and the glgC gene and the maize starch soluble synthase gene I, the fill length SSI. The N-terminal sequence is CVAELSREGPA

[0042]FIG. 10 shows plasmid pEXSCA with 6738 base pairs and promoter T7 and a Kanamycin gene and the glgC gene and the glgA gene

[0043]FIG. 11 shows plasmid pEXSC9a with 7240 base pairs and promoter T7 and ampicillin gene and the maize starch soluble synthase gene IIb-2 (Maize SS IIb-2), an N-terminally-truncated SSIIb. The N-terminal sequence is MNVVVVASECAP.

[0044]FIG. 12 shows plasmid pEXSWX with 6968 base pairs and promoter T7 and an ampicillin gene and the N-terminally-truncated maize WX (maize granular bound starch synthase). The N-terminal sequence for wx is ASAGMNVVFVGAEMA.

[0045]FIG. 13 shows plasmid pEXSWX2 with 6980 base pairs and promoter T7 and an ampicillin gene and the N-terminally-truncated maize WX termed as wx2. The N-terminus of wx2 is MNVVFVGAEMA.

[0046]FIG. 14 shows plasmid pEXSC9 with 7780 base pairs and promoter T7 and ampicillin gene and E. coli glgC gene and the maize starch soluble synthase geneIIb (Maize SS IIb).

[0047]FIG. 15 shows plasmid pEXSC10d with 7112 base pairs and promoter T7 and ampicillin gene, E. coli glgC gene and the N-terminally-truncated maize starch soluble synthase gene I termed as Maize SSI-3). The N-terminus of maize SSI-3 is MSIVFTGEASPYA.

[0048]FIG. 16 shows plasmid pEXS10 with 5300 base pairs and promoter T7 and ampicillin gene and the full length maize starch soluble synthase gene I termed as Maize SS 1.

[0049]FIG. 17 shows plasmid pEXS8 with 7259 base pairs and promoter T7 and ampicillin gene and the N-terminally-truncated maize starch synthase gene I termed as SSI-2. The N-terminal sequence is CVAELSRDLGLEPEG.

[0050]FIG. 18 shows plasmid pEXSCA1 with 5128 base pairs and promoter T7 and ampicillin gene and the gigA. pESCA1 is a 1551 bp SpeI-Sac I fragment containing glgA (from glgA in pBSK) subcloned into the Xba I-Sac I sites of p ET-23d which is commercially available from Novagen in Madision Wis. under catalog number 69748-1 and called ET-23d(+) DNA.

[0051]FIG. 19 shows the spectrum of the iodine glucan complex of the product produced by the host containing the glgC and glgA, and the pEXSC9, pEXSC3, pEXSC8, pEXSCwx the X-axis is listing nm and the Y axis is reading absorbance.

[0052]FIG. 20 shows the spectrum of the iodine glucan complex of the product produced by the host transformed with plasmids containing the glgC, the BEI, the BEII genes and glgA; glgC, the BEI, the BEII genes and maize SSI, SSI-2 and glgC, the BEI, the BEII genes and maize SSIIb, and glgC, the BEI, the BEII genes and maize SSIIa-2, and glgC, the BEI, the BEII genes, the X-axis is listing nm and the Y axis is reading absorbance.

[0053]FIG. 21 shows the product produced by the host in small bottles including the product from the host containing glgC, the BEI, the BEII genes and maize SS=s genes. Encoded as (C−I−II+8), glgC, the BEI, the BEII and maize SSI-2 genes and pEXSC10 encoded as (C-1-II+10), glgC, the BEI, the BEII and maize SSI genes and pEXSC9 encoded as (C-1-II+9),glgC, the BEI, the BEII and maize SSIIb genes and pEXSC3a encoded as (C−1−I+3 a), glgC, the BEI, the BEII and maize SSIIa-2 genes and pEXSCWX encoded as (C−1−II+WX),glgC, the BEI, the BEII and maize waxy genes and pEXSCA1 encoded as (C−T−II+A1), containing maize BEI, BEII and E. coli glgA genes, potato dextrin, waxy maize starch, corn amylopectin, rice starch, corn starch, pEXSC8.

[0054]FIG. 22 shows pExs-trc has 4178 base pairs with the trc promoter and the ampicillin gene. PEXS trc is pTrc99A-Nde1 which has been mutagenixed. (Nco I site in multiple cloning site of p Trc99A-NdeI is mutagenixed to Nde I using primers EXS63) AND EXS64.) pEXS-trc contains only one Nde I site and no Nco I sites. The following sites are not contained in pEXS-trc; Bgl II. Cla I, Nco I, Not I, Sac II, SnaB I, Spe I, and Xho I.

[0055]FIG. 23 shows pEXS-trc3 has 4129 base pairs with the trc promoter and the ampicillin gene in partial and the Kanamycin gene. The pEXS-trc3 is pEXS-tr1 cut with BgII (filled in)-Sca I and religated, deleting most of the Amp gene (304 nt from the 5″ end remain). The following sites are Not contained in p EXS-trc3: Apa I, Bgl II, Eco V, Nco I, Not l, SnaB I, and Spe I.

[0056]FIG. 24 shows the plasmid pEXS 102 having 7190 base pairs, adapted for plant transformation containing the maize 10 KD zein promoter, and maize adh I intron, the gene coding for the maize starch synthase I transit peptide, and the Waxy 2 gene and the nos terminator and the ampicillin gene.

[0057]FIG. 25 shows the plasmid pEXS 103 having 6607 base pairs, adapted for plant use containing the maize 10 KD zein promoter,the gene coding for the maize starch synthase I transit peptide and the Waxy 2 gene and the nos terminator and the ampicillin gene.

[0058]FIG. 26 shows the plasmid pEXS 101 having 6979 base pairs, adapted for plant use containing the maize 10 KD zein promoter, the gene coding for the maize starch synthase I transit peptide and the gig B gene and the nos terminator and the ampicillin gene.

[0059]FIG. 27 shows the plasmid pEXS 100 having 7557 base pairs, adapted for plant use containing the maize 10 KD zein promoter, and maize adh I intron, the gene coding for the maize starch synthase I transit peptide, and the glg B. gene and the nos terminator and the ampicillin gene.

[0060]FIG. 28 shows the plasmid pEXS 101 having 6273 base pairs. adapted for plant use containing the maize 10 KD zein promoter, the gene coding for the maize starch synthase I transit peptide, and the gig A gene and the nos terminator and the ampicillin gene.

[0061]FIG. 29 shows the plasmid pEXS 66 having 6001 base pairs, adapted for plant use containing the maize 10 KD zein promoter, the gene coding for the maize starch synthase I transit peptide, and the gig C₃ gene and the nos terminator and the ampicillin gene.

[0062]FIG. 30 shows the plasmid pEXS 65 having 6373 base pairs, adapted. for plant use containing the maize 10 KD zein promoter, the gene coding for the. maize starch synthase I transit peptide, and the maize waxy gene and the nos. terminator and the ampicillin gene.

[0063]FIG. 31 shows the plasmid pEXS 64 having 7073 base pairs, adapted for plant use containing the maize 10 KD zein promoter,the gene coding for the maize starch synthase I transit peptide, and the maize soluble starch synthase IIa gene and the nos terminator and the ampicillin gene.

[0064]FIG. 32 shows the plasmid pEXS 63 having 6473 base pairs, adapted for plant use containing the maize 10 KD zein promoter, the gene coding for the maize starch synthase I transit peptide, and the maize soluble starch synthase IIa gene and the nos terminator and the ampicillin gene.

[0065]FIG. 33 shows the plasmid pEXS 62 having 6773 base pairs, adapted for plant use containing the maize 10 KD zein promoter,the gene coding for the maize starch synthase I transit peptide, and the maize soluble starch synthase 1-2 gene and the nos terminator and the ampicillin gene

[0066]FIG. 34 shows the plasmid pEXS 61 having 7013 base pairs, adapted for plant use containing the maize 10 KD zein promoter, the gene coding for the maize starch synthase I transit peptide, and the maize soluble starch synthase IIb gene and the nos terminator and the ampicillin gene.

[0067]FIG. 35 shows the plasmid pEXS 59 having 6858 base pairs, adapted for plant use containing the maize 10 KD zein promoter, and maize adh I intron, the gene coding for the maize starch synthase I transit peptide, and the E. coli glgA gene and the nos terminator and the ampicillin gene

[0068]FIG. 36 shows the plasmid pEXS 58 having 7658 base pairs, adapted for plant use containing the maize 10 KD zein promoter, and maize adh I intron, the gene coding for the maize starch synthase I transit peptide, and the maize soluble starch synthase IIa gene and the nos terminator and the ampicillin gene.

[0069]FIG. 37 shows the plasmid pEXS 56 having 6586 base pairs, adapted for plant use containing the maize 10 KD zein promoter, and maize adh I intron, the gene coding for the maize starch synthase I transit peptide, and the gig C₃ gene and the nos terminator and the ampicillin gene.

[0070]FIG. 38 shows the plasmid pEXS 54 having 7658 base pairs, adapted for plant use containing the maize 10 KD zein promoter, and maize adh I intron, the gene coding for the maize starch synthase I transit peptide, and the Maize SS IIa gene and the nos terminator and the ampicillin gene.

[0071]FIG. 39 shows the plasmid pEXS 53 having 7058 base pairs, adapted for plant use containing the maize 10 KD zein promoter, and maize adh I intron, the gene coding for the maize starch synthase I transit peptide. and the maize starch soluble synthase IIa-2 gene and the nos terminator and the ampicillin gene.

[0072]FIG. 40 shows the plasmid pEXS 52 having 7358 base pairs, adapted for plant use containing the maize 10 KD zein promoter, and maize adh I intron. the gene coding for the maize starch synthase I transit peptide, and maize starch soluble synthase 1-2 gene and the nos terminator and the ampicillin gene.

[0073]FIG. 41 shows the plasmid pEXS 51 having 7398 base pairs, adapted for plant use containing the maize 10 KD zein promoter, and maize adh I intron, the gene coding for the maize starch synthase I transit peptide, and maize starch soluble synthase IIb gene and the nos terminator and the ampicillin gene.

[0074]FIG. 42 shows photograph of eleven products of altered starch produced with the present invention. The titled are encoded C−I−I=the glgC gene and the BEI and the BEII genes the following number or alternatively designation means pEXS-and the number. Thus C−I−II=the glgC gene and the BEI and the BEII and EXS-10 plasmid that contains the gene SSI, having the N-terminus shown in Table 1.

[0075]FIG. 43 shows the DNA sequence and the protein sequence for glgA having 1488 base pairs.

[0076]FIG. 44 shows the DNA sequence and the protein sequence for glgB having 2361 base pairs.

[0077]FIG. 45a shows the DNA sequence for Zea mays 10-kDa zein gene having 2562 base pairs.

[0078]FIG. 45b shows the DNA sequence for Zea mays 10-kDa zein portion of the gene used as the promoter in a number of the plasmids discussed herein

[0079]FIG. 46 shows the DNA sequence and the protein sequence for glgC3 (glgC₃) having 1328 base pairs containing two mutations P295D, K296E. This is a mutant of the wild type glgC gene.

[0080]FIG. 47 shows the DNA sequence and the protein sequence for glgC (glgC) having 1328 base pairs.

[0081]FIG. 48 shows the DNA sequence and the protein sequence for glgCwt (glgCwt) having 1328 base pairs. This is the glgC gene that is found in nature.

[0082]FIG. 49 shows the DNA sequence and the protein sequence for the maize waxy gene denoted wx herein.

[0083]FIG. 50 shows the DNA sequence and the protein sequence for the maize starch soluble synthase IIb encoding gene having 2423 base pairs.

[0084]FIG. 51 shows the DNA sequence and the protein sequence for the maize starch soluble synthase Ia.

[0085]FIG. 52 shows the DNA sequence and the protein sequence for the maize starch soluble synthase 1-2 having 1749 base pairs.

[0086]FIG. 53 shows the DNA sequence and the protein sequence for the maize branching enzyme II.

[0087]FIG. 54 shows the DNA sequence and the protein sequence for the maize branching enzyme I.

[0088]FIG. 55 shows the DNA sequence and the protein sequence (153) for the transit peptide portion of the maize starch soluble synthase I.

[0089]FIG. 56, PCR analysis of transgenic rice plants. The genomic DNA isolated from rice plants were PCR amplified using specific primers for the inserted gene. The specific bands were identified on 1% agarose gel compared with non-transgenic rice plant.

[0090]FIG. 57. Activity staining of starch synthase on renaturing SDS-PAGE gel with Iodine solution The positive staining of maize SSI-2 indicated the expression of maize SSI-2 in transgenic rice plants.

[0091]FIG. 58. SSI-1, SSI-2, and SSI-3 construct diagram. Three. forms of SSI were constructed in the pET expression system (see Methods). pExs10a encodes SSI-1, the fill length maize SSI (583 amino acids). pExs8 encodes a truncated SSI, SSI-2, with amino acids #8-52 deleted from the N-terminus of SSI-1. pExsld encodes the most truncated form of SSI, SSI-3, with the first 93 amino acids deleted from SSI-1. A depiction of the wax gene, encoding GBSS, is also included for comparison. The amino acid motif KS/TGGL, the putative binding site for ADPGlc, is indicated by the triangles. The KS/TGGL motif is located 18 amino acids from the N-terminus in GBSS, while the motif is 106 amino acids from the N-terminus in maize SSI. Drawing not to scale.

[0092]FIG. 59. SSIIa-1 and SSIIa-2 construct diagram. Two forms of SSIIa were constructed in the pET expression system. pExs3c encodes SSIIb-1, the putative full length maize SSIIb. N-terminal sequencing of SSIIa-1 revealed that the polypeptide chain started at amino acid #1, so the length of SSIIa-1 is 669 amino acids. pExs3a encodes a truncated form of SSIIa SSIIa-2, with the first 176 N-terminal amino acids deleted from SSIIa (493 amino acids total). A depiction of the waxy gene, encoding GBSS, is also included for comparison. The amino acid motif KTGGL, the putative binding site for ADPGlc, is indicated by the triangles. The KTGGL motif is located 18 amino acids from the N-terminus in GBSS, while the motif is 194 amino acids from the N-terminus in maize SSIIa

[0093]FIG. 60. SSIIb-1 and SSIIb-2 construct diagram. Two forms of SSIIb were constructed in the pET expression system (see Methods). pExs9 encodes SSIIb-1, the putative full length maize SSIIb. N-terminal sequencing of SSIIb-1 revealed that the polypeptide chain started at amino acid #1, so the length of SSIIb-1 is 637 amino acids. pExs9a encodes a truncated form of SSIIb, SSIIb-2, with the first 144 N-terminal amino acids deleted from SSIIb (492 amino acids total). A depiction of the waxy gene, encoding GBSS, is also included for comparison. The amino acid motif KTGGL, the putative binding site for ADPGlc, is indicated by the triangles. The KTGGL motif is located 18 amino acids from the N-terminus in GBSS, while the motif is 158 amino acids from the N-terminus in maize SSIIb.

[0094]FIG. 61. Temperature Curves for SSI enzymes. All assay components, except enzyme and [U-¹⁴]-ADPGlc, were mixed and then preincubated at each temperature for 3 minutes before addition of enzyme and ADPGlc. For all assays, the final concentration of [U-¹⁴C]-ADPGlc was 3 mM, while amylopectin was 6 mg/ml. Each point is an average of three separate determinations.

[0095]FIG. 62. Temperature Optima of SSIIa-1 and SSIIa-2. All assay components, except enzyme and [U-¹⁴C]-ADPGlc, were mixed and then preincubated at reach temperature for 3 minutes before addition of enzyme and ADPGlc. For assays in the presence of 0.5 M citrate, 5 mg/l ml amylopectin was used as primer. For assays without citrate, 10 mg/ml amylopectin was used. For all assays, the concentration of [U-¹⁴C]-ADPGlc was 3 mM. Each point is an average of three separate determinations.

[0096]FIG. 63. Temperature Optima of SSIIb-I and SSIIb-2. All assay components, except enzyme and [U-¹⁴C]ADPGlc, were mixed and then preincubated at each temperature for 3 minutes before addition of enzyme and ADPGlc. For all assays, the concentration of [U-¹⁴C]AFPGlc was 3 mM and the concentration of glycogen was 40 mg/ml. Each point is an average of three separate determinations.

PREFERRED EMBODIMENT—DESCRIPTION

[0097] Gene shall mean the entire gene sequence or any mutations or varieties of the codon that produce the desired activity in the host or alternatively the section or sections of the gene sequence necessary to produce the desired activity in the host. For example glgC gene shall mean glgC₁₆, glgC₃ and other mutants that produce the desired activity in the host. Starch synthase gene shall mean full length SS, N-terminally-truncated SS or mutated SS with starch synthase activity.

[0098] Glycogen like—shall mean polysaccharide material such as those produced as the main starch product by E. coli in its native state and by the hosts as taught in the above described paper by Hanping Guan.

[0099] Non Glycogen like—shall mean polysaccharide material which is plant like and is not produced as the main starch product by E. coli in its native state and by the hosts as taught in the above described paper by Hanping Guan.

[0100] Plant like starch—is non glycogen like.

[0101] Transformed gene—shall mean a gene that was somewhere in the lineage of the plant or bacteria introduced into the plant by means other than nature. Thus the progeny of a transformed host would continue to contain a transformed gene.

[0102] Transformed host—shall mean any organism containing one or more of the novel plasmids and/or a novel combination of starch synthetic genes discussed herein. Within this application a number of different protocols have been employed to designate the same gene or synthase. MSS#=maize soluble starch synthase, SS#will likewise mean starch synthase though not necessarily maize. STS#will also designate soluble starch synthase. GBSS=granule bound starch synthase. SBE#=starch branching enzyme, MBE=maize starch branching enzyme, MSBE#=maize starch branching enzyme, and BE#=starch branching. enzyme.

[0103] The present invention broadly encompasses transforming hosts such as bacteria or plants with plant starch synthetic genes that produce a non glycogen like material (a bacteria containing BEI and BEII from maize produces a glycogen like material). Starch bearing plants and organisms hereinafter are referred to as the host. One of the primary aspects of this invention is the generation of plant like starch from a bacterial host and the production of altered starch in a plant host. The present invention has been exemplified in both bacteria and in transformed rice plants. The host can contain though it is not a limitation, an unlimited supply of ADPG from the addition of the glgC gene (the bacterial gene) to the plant. Additionally the present invention encompasses plasmids that contain the maize genes and/or the bacterial genes in a construct adapted for use in a bacteria and constructs adapted for use in a plant. The plasmids in the plant construct preferably containing an active promoter recognized by the plant, a transit peptide, and the cleavage site that permits the protein to cleave from the transit peptide when crossing into the amyloplast in the plant. The plasmids used in the rice transformation specifically encompassed the maize 10 kd zein promoter, and the transit peptide from the maize SSI gene in the constructs adapted for plant use. The present invention also encompasses the plant producing the altered starch in the starch storage section of the plant or within the host cell and the altered starch itself. Additionally the present invention encompasses the combination of a number of starch genes in combination being active in a host such that the host produces differing non glycogen polysaccharides. Still further the present invention encompasses a method of making plant like starch in a bacterial host and the method of making altered plant like starch (altered in relationship to the type or amount of starch that the host makes without the constructs containing the genes), in a plant. Yet another object of the present invention is the addition of a gene that encodes for the substrate ADPG used to form starch.

[0104] The present invention encompasses a plasmid or combination of plasmids in the same host having a promoter adapted for use in a plant and a gene encoding for ADPGlc Pyrophosphoroylase, preferably a bacterial gene, and a gene encoding for starch synthase I or its mutant form. The present invention also encompasses the combination of a promoter adapted for use in a plant and optionally a gene encoding for ADPGlc Pyrophosphoroylase, preferably a bacterial gene, and a gene encoding for starch synthase I or its mutant form. and at least one gene encoding for branching enzyme transformed into a plant host.

[0105] The present invention encompasses a plasmid or combination of plasmids in the same host having a promoter adapted for use in a plant and a gene encoding for ADPGlc pyrophosphorylase, preferably a bacterial gene, and a gene encoding for starch synthase Iia or its mutant form. The present invention also encompasses the combination of a promoter adapted for use in a plant and optionally a gene encoding for ADPGlc pyrophosphorylase, and a gene encoding for starch synthase Iia or its mutant form, and at least one gene encoding for branching enzyme transformed in to a plant host.

[0106] The present invention encompasses a plasmid having a promoter adapted for use in a plant and a gene encoding for ADPGlc pyrophosphorylase, preferably a bacterial gene. and a gene encoding for starch synthase IIb and its mutant form. The present invention also encompasses the combination for a promoter adapted of use in a plant and an optional gene encoding for ADPGlc pyrophosphorylase, and a gene encoding for starch synthase IIb or its mutant form and at least one gene encoding for branching enzyme transformed in to a plant host.

[0107] The present invention encompasses a plasmid having a promoter adapted for use in a plant and a gene encoding for Pyrophosphoroylase, preferably a bacterial gene. and genes encoding for at least one of the following genes starch synthase I. starch synthase IIa starch synthase IIb, DUI. The present invention also encompasses the combination of a promoter adapted for use in a plant and a gene encoding for ADPGlc Pyrophosphoroylase, preferably a bacterial gene, and genes encoding for at least one of the following genes starch synthase 1, starch synthase IIa. starch synthase Iib and DUI, and at least one gene encoding for branching enzyme transformed in to a plant host.

[0108] The present invention encompasses a plasmid or combination of plasmids in the same host having a promoter adapted for use in a plant and a gene encoding for ADPGlc Pyrophosphoroylase, preferably a bacterial gene, and genes encoding for at least one of the following genes starch synthase I, starch synthase Ia, IIb and starch synthase III (DUI). The present invention also encompasses the combination of a promoter adapted for use in a plant and an optional gene encoding for ADPGlc pyrophosphorylase, preferably a bacterial gene, and genes encoding for at least one of the following genes starch synthase I, starch synthase Ia, IIb starch synthase III(DU1), and at least one gene encoding for branching enzyme. and at least one gene encoding for the debranching enzyme transformed in to a plant host.

[0109] The present invention encompasses a plasmid or combination of plasmids in the host having a promoter adapted for use in a bacteria or yeast and a gene encoding for ADPGlc Pyrophosphoroylase, preferably a bacterial gene, and a gene encoding for starch synthase I. The present invention also encompasses the combination of a promoter adapted for use in a bacteria or yeast and a gene encoding for ADPGlc Pyrophosphoroylase, preferably a bacterial gene, and a gene encoding for starch synthase I, and at least one gene encoding for branching enzyme transformed in to a bacteria or yeast host.

[0110] The present invention encompasses a plasmid or combination of plasmids in the host having a promoter adapted for use in a bacteria or yeast and a gene encoding for Pyrophosphoroylase, preferably a bacterial gene, and a gene encoding for starch synthase IIa. The present invention also encompasses the combination of a promoter adapted for use in a bacteria or yeast and optionally a gene encoding for ADPGlc Pyrophosphoroylase, preferably a bacterial gene, and a gene encoding for starch synthase IIa, and at least one gene encoding for branching enzyme transformed in to a bacteria or yeast host.

[0111] The present invention encompasses a plasmid or combination of plasmids in the same host having a promoter adapted for use in a bacteria or yeast, and a maize gene encoding for starch synthase III(DU1). The present invention also encompasses the combination of a promoter adapted for use in a bacteria or yeast and an optional gene encoding for ADPGlc Pyrophosphoroylase, preferably a bacterial gene, and a gene encoding for starch synthase III, and at least one gene encoding for branching enzyme transformed in to a bacteria or yeast host.

[0112] The present invention encompasses a plasmid or combination of plasmids in the same host having a promoter adapted for use in bacteria or in yeast and a gene, and genes encoding for at least one of the following genes starch synthase I, starch synthase IIa, IIb, starch synthase III(DU1). The present invention also encompasses the combination of a promoter adapted for use in bacteria or in yeast and a gene encoding for ADPGlc Pyrophosphoroylase, preferably a bacterial gene, and genes encoding for at least one of the following genes starch synthase I, starch synthase IIa,IIb, starch synthase III, and at least one gene encoding for branching enzyme transformed in to bacteria or into yeast hosts.

[0113] The present invention encompasses a plasmid or combination of plasmids in the same host having a promoter adapted for use in bacteria or in yeast and a gene encoding for ADPGlc Pyrophosphoroylase, preferably a bacterial gene, and genes encoding for at least one of the following genes starch synthase 1, starch synthase IIa,IIb, starch synthase III The present invention also encompasses the combination of a promoter adapted for use in bacteria or in yeast and a gene encoding for Pyrophosphoroylase, preferably a bacterial gene, and genes encoding for at least one of the following genes starch synthase I, starch synthase IIa IIb. starch synthase III(DU1), and at least one gene encoding for branching enzyme, and at least one gene encoding for the debranching enzyme transformed in to a bacteria or into a yeast host.

[0114] The present invention encompasses the truncated versions of the SSI and the SSII and the SSIII genes that still provide protein that is sufficient to make the polysaccharide. By transforming different combinations of SS and SBE into E. coli HPG204(DE3) or G6MD3 defective in GS and GBE, we obtained the first evidence that maize SSI, SSII and SSIII have different specificities in the size of glucans synthesized see fig one. Herein, we present the model system to produce differing polysaccharides from hosts with SS and SBE in E. coli by metabolic engineering. We also demonstrated that the truncated forms of SS had different Vmax, temperature stability and kinetic properties (Table, Fig).

[0115] We also demonstrated that transformation of starch synthase and/or branching enzyme in E coli resulted in production of polysaccharides differing in size and structure. These polysaccharides can be used in food and nonfood industries to replace and/or complement starch functionalities. A large amount of these polysaccharides can be produced with fermentation technology.

[0116] Starch biosynthesis in higher plants and glycogen biosynthesis in E. coli have similar reactions which use adenosine diphosphate glucose (ADPGlc) as a substrate. This similarity allows us to use plant starch synthase (SS) and starch branching enzyme (SBE) to complement the functions of glycogen synthase (GS) and glycogen branching enzyme (GBE) in E. coli G6MD3, which is deficient in GS and GBE. Transformation of E. coli glgC gene and maize starch synthase gene in E. coli G6MD3 produced linear a 1,4 glucan similar to amylose. coexpression of the glgC, maize starch synthase and maize branching enzyme produced branched polysaccharides. However, distinct properties of plant starch branching enzyme and starch synthase make it possible to synthesize different polysaccharides in E. coli. While maize SSI preferentially synthesis short chains (dp 6-15), SSII and SSIII preferentially transferred long chains (dp>24) and intermediate chains (dp 16-24) respectively. Transformation of different maize starch synthases, E. coli glycogen synthase (glgA) and/or maize branching enzymes into E. coli HPG96 or E. coli G6MD3 resulted in the synthesis of different sizes of polysaccharide with DP 500-4000.These polysaccharides synthesized in E coli by maize SS have different physical-chemical properties than polysaccharides synthesized in natural organisms including starch from plant sources and glycogen from animals. The polysaccharide can be used in food and nonfood industries to replace and/or complement starch functionalities. A large amount of these polysaccharide can be produced by fermentation technology. The following materials were employed in the construction of the present invention some of the starting material are commercially available from Novacen in Madision Wis. ET-23d(+) DNA under catalog number 69748-1 and BL21(DE3) under catalog number 69387-1; ET-2 la(+) DNA under catalog number 697401.

[0117] Plant Hosts

[0118] The following plasmids have been transformed into rice plants Transgenic 1, MSTSIA(pExs52) and glgC₃ (pExs66), MSTSIIa and glgC₃ (pExs53 and pExs56). The second group of rice transformatns contain MSTSIIc and glgC₃ (pExs54 and pExs56).The third group, of transformation: transgenic 5 MSTSIII and glgC₃ (pExs 61 and pExs 66); transgenic 6 Mwx glgC₃ p Exs65 and pExs66). Generally see FIGS. 2541 for plasmid maps and FIGS. 43-55 for sequences used in the plasmid. Additionally, glgA and glgB and glgC were combined and transformed into rice. This is combining the rice plants starch pathway with the gene encoding for ADPG and the genes encoding for at least one of the following enzymes, SSI, SSII, SSIII, Debranching enzymes, BEI, BEII, GBSS (wx).

[0119] These plasmids could have been transformed into other cereals such as corn, wheat, barley, oats, sorghum, milo in substantially the plasmid that is shown in the figures for the plant host. The promoter could be the waxy gene which is published, other additional zein promoters are known and could be used as the promoter. The promoter used herein is described in FIGS. 45a and 45b.

[0120] Additionally these plasmid with little additional work could be transformed into dicots such as potatoes, sweet potato, taro, yam, lotus cassaya, peanuts, peas., soybean, beans, chickpeas. The promoter could be selected to target the starch storage area of the particular dicots (some are roots some are tubers). Various method. of transforming monocots and dicots are known in the industry and the method of transforming the genes is not critical to the present invention. The plasmid can be introduced into Agrobacterium tumefaciens by the, freeze-thaw method of An et al.(1988) Binary vectors. In Plant Molecular Biology Manual A3. S. B. Gelvin and R. A. Schilperoot.eds (Dordrecht, The Netherlands: Kluwer Academic Publishers), pp. 1-19. Preparation of Agrobacteruim inoculum carrying the construct and inoculation of plant material, regeneration of shoots, and rooting of shoots are described in Edwards et al. (1995). Biochemical and molecular characterization of a novel starch synthase from potatoes. Plant J. 8, 283-294. Additionally promoters for different dicots are known particularly 15sCaMV and Monsanto has also published a promoter that is useful in potatoes called a patatin promoter.

[0121] A number of monocots are also starch bearing plants but until about a decade ago monocots were difficult to develop transformants. The most prominent methods of transformation presently used in monocots is the gunning of micro projectiles into the plants or using Agrobacterium and subsequent regeneration of the plants from the transformed materials. Various tissues and cells can now be transformed with plasmids into monocot hosts. In fact there are teaching from at least five ago on methods of transforming not only callus, but also cotyledons. The methods of transforming plants and selecting for the transformants with either selectable or screen able markers are also well known. The use of the marker in the same, plasmid and the use of the markers in a separate plasmid that is co transformed into the host are well known in the art by those of ordinary skill in the art. The biotechnology methods of forming plasmids and transforming plants are listed in the book entitled AShort Protocols In Molecular Biology@, 3rd ed. Published in 1995 by JOHN WILEY & Sons, Inc. Additionally, methods of transforming with the gun and with protoplasts are taught in a number of issued patents to Dekalb and Agracetus and Ciba.

PREFERRED EMBODIMENT—OPERATION EXAMPLE 1 Construction of the E. coli Expression Vector

[0122] The expression vector pExs2 was derived from pET-23d (Novagen) and pGP1-2 (15). The expression vectors pExs-trc and pExs-trc3 were derived from pTrc99a (Pharmacia) and pGP1-2. The BglII/PstI fragment (2192 bp) containing the pBR322 origin of replication was deleted from pET-23d and replaced with the BamHI/PstI fragment (3 kb) containing origin p15A and kanamycin resistance gene from pGPI-2. This process generated plasmid pEXS1 containing both ampicillin and kanamycin resistance genes. The ampicillin resistance gene was inactivated by deletion of the ScaI BglI fragment (360 bp, BglI end was filled in and blunt-end ligated with ScaI end). Inactivation of the ampicillin resistance gene in pEXS I generated the expression plasmid pEXS2, containing the T7 promoter, T7 terminator; kanamycin resistance gene and p15A origin of replication. Plasmid pTrc99a was digested with Nde1 filled in with kelnow fragment and blunt-end ligated to remove NdeI site. A NdeI site was introduced at the NcoI site by mutagenesis to generate plasmid pExs-trc. The BglI and PvuII fragment (2.48 kb) in pExs-trc containing the pBR322 origin of replication was replaced by BglII/BamHI (filled in with Klenow fragment) fragment (3 kb) containing origin p15A and kanamycin resistance gene from pGP1-2 to generate pExs-trc2. The ampicillin resistance gene was inactivated by deletion of the ScaI/BglI fragment (360 bp, BglI end was filled in and blunt-end ligated with ScaI end). Inactivation of the ampicillin resistance gene in pExs-trc2 generated the expression plasmid pExs-trc3.

[0123] Construction of expression plasmids for maize SS. For expression of maize SS in E. coli, The PCR method was used to modify the N-terminus of maize SS using the following nucleiotides: primer Exs4 (5′-CAAGAATGCTGCGGGAGTC-3′), primer Exs23 (5′-AAGTCGACATATGTGCGTCGCGGAGCTGAGCAG-3′), primer Exs 57 (5′-GGGCCCCATATGAGCATTGTCTTTGTAACCGG-3′), primer Exs I (5′-CTCGGGCCCATATGGGGGAGAATGTTATGAA-3′), primer Exs2 (5′-GAGGCATCAATGAACACAAAGTCG-3′), Primer Exs33 (5′-GAAGGGCCCCATATGGCTGAGGCTGAGGCCGGGGGCAAG-3′), primer Exs 16 (5′-TTOGATCCATATGGGAGCTGCGGTTGCATTGGG-3′) and primer Exs17 (5′-CCTGCGGGCTCTGGCTTCACC), primer Exs 55 (5′-TTGGATCCATATGAACGTCGTCGTGGTGGCTTC-3′), primer 56 (5′-GCATACCATGGAACCTCAACAGC-3′), primer 53 (5′-GGTACCATATGAACGTCGTCTTCGGCG-3′), primer Exs 54 (5′-GACAGGCCCGTAGATCTTCTCC-3′), primer Exs-wx (5′-TTGGTACCATATGGCCAGCGCCGCCGGCATGAACG-3′). Primer Exs 4 paired respectively with primer Exs23 and Exs 57 was to modify the N-terminus of maize SSS1 gene to generate pExs-10 and pExs-1d. Primer Exs2 paired individually with primer Exs33 and Exs1 was to modify the N-terminus of maize SSSII to generate pExs3c and pExs3a. Primer Exs17 paired individually with primer Exs16 and Exs55 was to modify the N-terminus of maize-SSSIII to generate pExs-9 and pExs-9a. Primer ExsS4 paired individually with primer Exs-wx and Exs53 was used to modify the N-terminus of maize: GBSS to generate pExs-wx and pExs-wx2. The modified N-terminus was recombined with the rest of the SS gene in pBluescript SK plasmid. The reconstructed of maize SS was subcloned from pBluescript SK to the NdeI/NotI sites of the expression vector pET-21a (Novagen), pExs-trc. pExs-trc3 (maps are attached, Table I shows the N-terminal sequence of SSS).

EXAMPLE 2

[0124] Construction of Expression Plasmids, for E. coli ADPGlc Pyrophosphorylase, BE and Maize SBE.

[0125]E. coli glgB gene was excised from plasmid pOP12 (16). The BstX1 (filled in)/HindIII fragment containing the glgB ribosome binding site and the full length glgB gene was cloned at the SmaI site of pBluescriptSK− (Stratagene). The glgB gene in pBluscriptSK− was subsequently cloned into pEXS2 at the XbaI/SalI sites to generate plasmid pEXSB. Primer G (5′-GAAGATCTGGCAGGGACCTGCACAC-3′) and primer H (5′-GGACTAGTGCATTATCGCTCCTGTTTAT-3′) were used to PCR the E coli glgC gene coding for ADPGlc pyrophosphorylase from plasmid pOP12. A BglII site and a SpeI site introduced by PCR to the N-terminal and. C-terminal site respectively, were used to clone the PCR product into pBluscript SK− at the BamHI and SpeI sites. The glgC gene including its own ribosome binding site was subcloned into expression plasmid pEXS2 at the XbaI (filled in with Klenow fragment) and NotI site to generate plasmid pEXSc. The genes coding for mature maize SBEI and SBEII along with a ribosome binding site were subcloned from plasmids pET-23d-SBEI and pET-23d-SBEII into the plasmid pEXSc at the SpeI site to form the plasmids pEXSc-SBEI and pEXSc-SBEII. The gene coding for mature maize SBEII including a ribosome binding site was cloned into pEXSc-SBEI at the XbaI/NotI sites to form plasmid pEXSc-SBEI-SBEII. E. coli glgc gene and genes encoding maize SBEI and SBEII were also cloned in plasmid pExs-trc and pExs-trc3 respectively and together as described for pExs2.

EXAMPLE 3 Isolation of E. coli HPG204 Deficient in GBE and GS Activities

[0126] Homologous recombination was used for the strain construction. This was done according to the method described by Hamilton et al (Journal of Bacteriology, 1989, 171:4617-4622.) A temperature-sensitive pSC10 replicon was used to facilitate the selection. The gene coding for spectinomycin adenyltransferase was inserted at PvuII sites in plasmid pOP12 to form plasmid HPG9 which has spectinomycin resistance and has C-terminus of glgB gene and N-terminus of glgA gene deleted. The DNA fragment B=SA=with Spectinomycin resistant gene inserted between partial truncated glgB and gigA was subcloned into plasmid pMAK705 at XbaI site containing temperature sensitive replicon (Hamilton et al. Journal of Bacteriology, 1989, 171:46174622.) to form Plasmid pMak705B=SA=. Plasmid pMak705B=SA=was transformed into TG1 cell. After the transformed cell was cultured in 3 mL LB with 100. mg/mL Spectinomycin at room temperature overnight, the cells were plated on LB agar plate containing 100 mg/mL spectinomycin and incubated at 44° C. overnight. Single colonies were inoculated on LB agar plate containing 100 mg/mL spectinomycin and 0.2% glucose and incubated at 44° C. and at 37° C. overnight. The colonies at 37° C. were stained with iodine. The colony with negative staining was selected and grown in 100 mL LB at 37° C. overnight. The cells were harvested and homogenized in an extraction buffer for assaying glycogen synthase and branching enzyme activities. The cell lacking glgA and glgB activities was named as HPG204 [F=traD36 LacI^(q)(glgBXCA) D(LacZ)M15proA⁺B⁻/SupED (hsdM-mcrB)5(r_(k) ⁻m_(k) ⁻McrB)thiD(lac-proAB),Spectinomycim^(R), Chloramphenicol^(R)]. The IDE3 lysogenization kit fromNovagen was used for site specific integration of IDE3 prophage into E. coli HPPG204 to form E. coli HPG204(DE3)[was DThe lysate was prepared with P1 vir and its transduction into E. coli BL21 (DE3) [F=traD36 Lacl^(q) D(glgBXCA) D(LacZ)M15proA⁺B⁺/SupED (hsdM-mcrB)5(r_(k) ⁻m_(k) ⁻McrB⁻)thiD(lac-proAB),spectinomycin^(R), Chloramphenicol^(R)]

EXAMPLE 4 Expression of Maize SS and SBE in E. coli

[0127] Plasmid pExs-9 and pExs-trc3 has kanamycin resistance and p15A origin of replication. It is compatible with plasmid pET21a, pExs-trc, pTrc99A containing pBR322 origin. Expression plasmids pExs-2 and pET-21a were used to express SS and SBE in E. coli HPG204(DE3). Expression plasmids pExs-trc and pExs-trc3 were used for expression in E. coli G6MD3. This made it possible to transform different combinations of maize SS and SBE in E. coli HPG204(DE3), or G6MD3 which is deficient in GS and GBE activity. An overnight culture of cells transformed with maize SS and SBE was diluted 1:20 (v/v) in fresh LB containing 0.2% glucose, 100 mg/mL ampicillin and 50 mg/mL kanamycin. The cells were grown at 37° C. for about 2 h to A600nm=0.6 before the expression of maize SBE and/or SS was induced by adding isopropyl b-D-thiogalactoside to 0.5 mM. Following growth at 25° C. for 4 h, the cells were harvested in a refrigerated centrifuge.

EXAMPLE 5 Isolation of Highly Branched a-glucan from E. coli

[0128] Cell pellet (30 g) was resuspended and lysed by sonication in 150 mL 50 mM tris-acetate buffer (pH 7.5) containing 10 mM EDTA and 5 mM DTT. After a fraction of the homogenate was saved for assaying the STS and SBE activities, the homogenate was centrifuged at 20,000 g for 50 min at 4° C. After collecting the supernatant, the pellet was resuspended in 150 nm water and boiled for 15 min with occasional stirring. The resuspension was centrifuged at 20,000 g for 30 at room temperature. After collecting the supernatant. the pellet was washed again with 100 mL water as above. 0.1 volumes of 50% Trichloric acid (TCA) were added to the pooled fractions. After storing on ice for 30 min, the precipitate was spun down at 15,000 g for 20 min, then washed with 30 mL 5% TCA and centrifuged as above. The supernatant and wash were pooled and one volume of absolute ethanol was added. After storing on ice for 30 min, the polysaccharide was collected by centrifuging at 15,000 g for 15 min. The polysaccharide was redissolved in water and precipitated with ethanol. This step was repeated twice. The pellet was washed with methanol twice, acetone twice and dried over silica gel at room temperature.

EXAMPLE 6 Isolation of Linear a 1,4 Polysaccharide from E. coli

[0129] Resuspend 50 grams of cell pellet in 250 mL of 50 mM Tris acetate buffer, pH 7.5, containing 10 mM EDTA and 5 mM DTT. Sonicate for 3 minutes (45 seconds/time, output #8, repeat 4 times with 30 seconds interval). The homogenate is centrifuged at 12,000 rpm (SA1500) for 50 minutes. The supernatant is checked with iodine staining and discarded. (Same 1 mL homogenate and; 1 mL supernatant for enzyme assay. The pellet is resuspended & extracted in 100 mL DMSO. Extract the polysaccharide by heating and stirring in boiling water bath for 15 min. Let it cool down to below 40° C. and centrifuge at 12.000 rpm for 30 min at room temperature. The supernatant is pooled. The pellet is extracted two more times with 100 mL DMSO. Equal volume of absolute ethanol is added into the pooled supernatant, mixed and stored on ice for 30. minutes. Centrifuge at 12,000 rpm for 30 min at 4° C. The pellet is redissolved in 20 mL DMSO by heating in boiling water bath. 80 mL water is added and mixed well. After adding. 10 mL butanol to the solution, the solution is mixed and stored at 0° C. for one hr (mix once a while). Centrifuge at 12000 rpm for 30 min at 40C. Repeat the step once. The pellet is redissolved in 90 mL hot water by heating in boiling water bath. Insoluble materials are immediately removed by centrifugation at room temperature. Add 10 mL butanol to the supernatant, stay at 0° C. for 1 hr and centrifuge at 12000 rpm for 30 min at 4° C. Repeat the step once. The amylose precipitate is redissolved in 90 mL hot water by heating, and 110 mL butanol are added to the solution. After storing at 40° C. on ice for one hour, it is centrifuged at 4° C. for 30 min. Repeat the step once. The pellet is redissolved in 100 mL 10% butanol by heating. The amylose is stored at 0° C. and precipitated by centrifuging at 12000 rpm for 30 min at 4° C. The pellet is washed with 25 mL methanol 3 times and with acetone once. Dry over silica gel.

EXAMPLE 7 Enzyme Assays

[0130] 5 mL of supernatant were used to assay STS and SBE activities as previously described (Press) with minor modification. The reaction mixture for STS contained 100 mM Bicine buffer, 10 mg/mL glycogen, 0.5 mg/ml BSA, 0.5 M sodium citrate, 25 mM potassium acetate, 10 mM GSH, 3 mM [14C]ADPGlc (500 dpm/nmol) and enzyme in a final volume of 0.1 mL. The reaction was carried out at 25° C. for 15 min and terminated by boiling for 2 min. The unincorporated [14C]ADPGlc was separated with Dowex anion exchange column (200-400 mesh, Sigma Chemical Co.). One unit of activity is defined as 1 mmol Glc incorporated into the a-glucan per min at 25° C. SBE activity was determined by phosphorylase stimulation assay. One unit of activity is defined as 1 mmol Glc incorporated into the a-glucan per min at 30° C.

Example 8 Enzyme Purification

[0131] For the recombinant SS purification, the cell pellet was resuspended in sonication buffer (50 mM Tris-acetate, pH 7.5, 10 mM EDTA, and 5 mM DTT; 7 ml buffer per gram of cell mass), and cells were lysed using a Fisher 550 Sonic Dismembrator with 5×1 min. bursts with 30 sec. intervals. The homogenate was centrifuged at 9600 g for 30 minutes. SSI in the supernatant was then precipitated by slowly adding neutralized saturated ammonium sulfate to 40% saturation. After stirring on ice for an additional 50 minutes, proteins were collected by centrifugation at 12700 g for 45 minutes. The protein pellet was then redissolved in buffer A (50 mM Tris, pH 7.5, 1 mM EDTA, and 5 mM DTT) containing 0.1 M KCl and dialyzed against the same buffer, with one change of buffer. After dialysis, the sample was centrifuged at 13000 g for 20 minutes to remove insoluble materials. The resulting supernatant was loaded onto an amylose affinity column. pre-equilibrated with dialysis buffer, and the flow through. was collected. The column was washed with 10 column volumes of buffer A containing 0.1 M KCl, and then with buffer A containing 0.5 M KCl and 0.5 M maltose, collecting fractions during both washes. The active fractions were pooled and dialyzed overnight against buffer A, with one change of buffer. The next day, the amylose column sample was filtered and applied to a mono Q 5/5 FPLC column (Pharmacia). After washing with buffer A, a 20 ml 0-0.4 M KCl gradient was employed. The active fractions were electrophoresed on an 8% SDS-PAGE gel (31) to determine the purity of SSI in those fractions; the fractions which were apparently homogeneous were pooled and concentrated using a Centricon-30 spin column (Amicon). TABLE 1 Expression of maize starch synthases in Escherichia coli BL21 (DE3). Specific Maize starch synthase Protein Activities Plasmids N-terminus genes (mg/mL) (units/mg Protein) pET21a Native plasmid 1.8 0.009 pEXS-3a SSIIa-2 GENVMNVIVV 2.8 0.069 pEXS-3c SSIIa AEAEAGGKD 2.8 0.28 pEXS-1d SS1-3 MSIVFVTGEA 3.0 0.23 pEXS-8 SSI-2 GDLGLEPEG 1.9 0.097 pExs-10 SSI CVAELSREG 1.2 0.043 pEXS-9 SSIIb GSVGAALRSY 1.8 0.515 pEXS-9a SSIIb-2 MNVVVVASEC 2.6 0.36 pEXS-wx GBSS (waxy) ASAGMNVVFV 2 0.033 pEXS-wx2 GBSS(2) MNVVFVGAEM 2.2 0.32

[0132] TABLE 2 Properties of polysaccharides synthesized in E. coli. Protein STS activity BE activity Imax Yield Plasmid (Mg/mL) (u/mg protein) (u/mg protein) (nm) DP CL (mg dry wt/g wet cell) pExsCA 580 700 10.6 3.3 pExsC-9 585 1007 35.8 4.1 pExsC-3a 13.3 .0015 600 983 53 1.0 pExsC-8 12.6 .0032 580 435 31.8 7.4 pExsC-wx 15.2 0.002 600 836 15.6 9.1 pExsC-I-II + pExs9 7.84 0.08 4.71 480 2333 19 30 pExsC-I-II + pExs3a 13.61 0.011 1.56 530 3616 22 36 pExsC-I-II + pExs8 11.95 0.042 3.33 525 1689 17.5 131 pExsC-I-II + pExs10 8.9 .0094 3.65 500 3174 16.6 24.5 pExsC-I-II + pExswx 11.7 .007 5.4 450 2970 14.8 33.8 pExsC-I-II + pExsA1 11 0.13 4.48 475 3940 14 28.9

[0133] TABLE 3 Properties listed by degree of DP of polysaccharides synthesized In E. coli. Protein STS activity BE activity Imax Yield Plasmid (Mg/mL) (u/mg protein) (u/mg protein) (nm) DP CL (mg dry wt/g wet cell) pExsC-I-II + pExsA1 11 0.13 4.48 475 3940 14 28.9 pExsC-I-II + pExs3a 13.61 0.011 1.56 530 3616 22 36 pExsC-I-II + pExs 108.9 .0094 3.65 500 3174 16.6 24.5 pExsC-I-II + pExswx 11.7 .007 5.4 450 2970 14.8 33.8 pExsC-I-II + pExs9 7.84 0.08 4.71 480 2333 19 30 pExsC-I-II + pExs8 11.95 0.042 3.33 525 1689 17.5 131 pExsC-9 585 1007 35.8 4.1 pExsC-3a 13.3 .0015 600 983 53 1.0 pExsC-wx 15.2 0.002 600 836 15.6 9.1 pExsCA 580 700 10.6 3.3 pExsC-8 12.6 .0032 580 435 31.8 7.4

[0134] TABLE 4 Properties listed by degree of λmax of polysaccharides synthesized in E. coli. Protein STS activity BE activity λmax Yield Plasmid (Mg/mL) (u/mg protein) (u/mg protein) (nm) DP CL (mg dry wt/g wet cell) pExsC-3a 13.3 .0015 600 983 53 1.0 pExsC-wx 15.2 0.002 600 836 15.6 9.1 pExsC-9 585 1007 35.8 4.1 pExsCA 580 700 10.6 3.3 pExsC-8 12.6 .0032 580 435 31.8 7.4 pExsC-I-II + pExs3a 13.61 0.011 1.56 530 3616 22 36 pExsC-I-II + pExs8 11.95 0.042 3.33 525 1689 17.5 131 pExsC-I-II + pExs10 8.9 .0094 3.65 500 3174 16.6 24.5 pExsC-I-II + pExs9 7.84 0.08 4.71 480 2333 19 30 pExsC-I-II + pExsA1 11 0.13 4.48 475 3940 14 28.9 pExsC-I-II + pExswx 11.7 .007 5.4 450 2970 14.8 33.8

[0135] TABLE 5 Properties listed by degree of CL of polysaccharides synthesized in E. coli. Protein STS activity BE activity λmax Yield Plasmid (Mg/mL) (u/mg protein) (u/mg protein) (nm) DP CL (mg dry wt/g wet cell) pExsC-3a 13.3 .0015 600 983 53 1.0 pExsC-9 585 1007 35.8 4.1 pExsC-8 12.6 .0032 580 435 31.8 7.4 pExsC-I-II + pExs3a 13.61 0.011 1.56 530 3616 22 36 pExsC-I-II + pExs9 7.84 0.08 4.71 480 2333 19 30 pExsC-I-II + pExs8 11.95 0.042 3.33 525 1689 17.5 131 pExsC-I-II + pExs10 8.9 .0094 3.65 500 3174 16.6 24.5 pExsC-wx 15.2 0.002 600 836 15.6 9.1 pExsC-I-II + pExswx 11.7 .007 5.4 450 2970 14.8 33.8 pExsC-I-II + pExsA1 11 0.13 4.48 475 3940 14 28.9 pExsCA 580 700 10.6 3.3

[0136] TABLE 6 Purification Tables for SSI-1, SSI-2, and SSI-3. volume total mg activity total purification (ml) protein U/mg Units (fold) SSI-1 Homogenate 630 4347 0.018 76.2 1 Supernatant 570 2622 0.020 53.0 1.1 0-40% (NH₄)₂SO₄ 48 494 0.058 28.7 3.2 Amylose column 17 2.6 5.03 11.3 279 monoQ column 0.27 0.26 12.2 3.2 677 SSI-2 Homogenate 380 2797 0.0356 99.6 1 Supernatant 320 2118 0.0340 72.0 1 0-40% (NH₄)₂SO₄ 48 466 0.133 61.8 3.7 Amylose column 17.5 1.2 22.6 26.5 634 monoQ column 1.0 0.325 17.2 5.6 483 SSI-3 Homogenate 1300 16770 0.23 3900 1 Supernatant 1100 9790 0.31 3080 1.3 0-40% (NH₄)₂SO₄ 237 2204 1.5 3294 6.5 Amylose column 63 30 22.4 668 97 monoQ column 3.6 3.1 30.5 93 132

[0137] TABLE 7 Primer Kinetics for SSI enzymes SSI-3 SSI-2 SSI-1 Amylopectin +citrate K_(m)  240 ± 45   230 ± 50   150 ± 40  V_(max) 26.3 ± 0.5 33.4 ± 2.1 22.5 ± 0.6 −citrate K_(m)  230 ± 60    68 ± 3    120 ± 20  V_(max) 13.2 ± 0.3  9.94 ± 0.18  7.62 ± 0.99 Glycogen +citrate ^(a)V_(max) 43.4 ± 2.5 45.6 ± 3.3 39.0 ± 2.2 −citrate ^(a)V_(max) 41.4 ± 2.9 45.5 ± 1.5 26.1 ± 1.4

[0138] TABLE 8 ADPGlc Kinetics for STSI enzymes. STSI-3 STSI-2 STSI-1 +citrate K_(m) 0.33 ± 0.07 0.32 ± 0.02 0.18 ± 0.02 V_(m) 26.4 ± 1.4  32.6 ± 0.8  18.0 ± 0.5  −citrate K_(m) 0.62 ± 0.04 0.25 ± 0.04 0.24 ± 0.02 V_(m) 14.7 ± 1.3  11.7 ± 0.7  6.38 ± 0.88

[0139] TABLE 9 Purification Tables for SSIIa enzymes. volume total mg activity total purification (ml) protein U/mg Units (fold) SSIIa-2 Supernatant 300 1620 0.0216 34.8 1 0-40% (NH₄)₂SO₄ 53 419 0.0606 25.4 2.8 Amylose column 20 9.3 0.991 9.3 45.9 monoQ column 0.9 0.94 4.81 4.5 222 SSIIa-1 Supernatant 335 2613 0.28 737 1 0-40% (NH₄)₂SO₄ 47 427 0.96 409 3.4 Amylose column 25 11.5 8.04 92 28.7 monoQ column 1.0 4.8 9.10 44 32.5

[0140] TABLE 10 Primer Kinetics for SSIIa enzymes. SSIIa-2 SSIIa-1 Amylopectin +citrate 27° C. K_(m) 153 ± 22 182 ± 38 V_(max)  7.82 ± 0.63 24.1 ± 0.5 37° C. K_(m) 133 ± 18 153 ± 64 V_(max) 15.4 ± 0.6 41.1 ± 0.2 −citrate 27° C. K_(m)  234 ± 3.0 404 ± 33 V_(max)  4.31 ± 0.32 10.5 ± 0.3 37° C. K_(m) 1350 ± 220 −NA* V_(max)  7.84 ± 0.25 −NA* Glycogen +citrate 27° C. K_(m) 50.7 ± 3.8 162 ± 17 V_(max)  5.53 ± 0.44 14.2 ± 0.7 37° C. K_(m) 76.9 ± 7.8 350 ± 11 V_(max) 11.3 ± 0.7 31.6 ± 0.8

[0141] TABLE 11 ADPGlc Kinetics for SSIIa enzymes. with amylopectin as primer SSIIa-2 SSIIa-1 +citrate 27° C. K_(m) 0.17 ± 0.04 0.48 ± 0.09 V_(max) 4.83 ± 0.42 23.0 ± 2.5  37° C. K_(m) 0.28 ± 0.01 0.83 ± 0.08 V_(max) 11.4 ± 0.6  49.1 ± 2.6  −citrate 27° C. K_(m) 0.27 ± 0.02 0.46 ± 0.06 V_(max) 4.87 ± 0.25 12.1 ± 0.8  37° C. K_(m)  0.28 ± 0.005 −NA* V_(max) 7.86 ± 0.53 −NA* with glycogen as primer with glycogen SSIIa-2 SSIIa-1 +citrate 27° C. K_(m) 0.16 ± 0.03 0.19 ± 0.02 V_(max) 4.41 ± 0.21 17.1 ± 0.7  37° C. K_(m) 0.15 ± 0.03 0.37 ± 0.04 V_(max) 7.60 ± 0.94 40.1 ± 1.7 

[0142] TABLE 12 Purification Tables for SSIIb-2 and SSIIb-1. volume total mg activity total purification (ml) protein U/mg Units fold SSIIb-2 Supernatant 890 9256 0.48 4450 1 0-40% (NH₄)₂SO₄ 190 2660 1.24 3306 2.6 Amylose column 13 31.2 50.6 1573 105 monoQ column 6.6 16.3 56.8 939 118 SSIIb-2 Supernatant 365 2336 0.64 1533 1 0-40% (NH₄)₂SO₄ 56 436 2.35 1030 3.7 Amylose column 80 10.4 50.2 521 78 monoQ column 0.6 0.28 60.6 17.6 94

[0143] TABLE 13 Kinetics for SSIIb enzymes. SSIIb-2 SSIIb-1 ADPGLc Kinetics with glycogen K_(m) 0.32 ± 0.04 0.71 ± 0.01 V_(max)  130 ± 6   76.8 ± 3.2  with amylopectin K_(m) 0.32 ± 0.03 0.40 ± 0.02 V_(max) 90.9 ± 4.2  72.8 ± 2.8  Primer Kinetics glycogen K_(m) 0.36 ± 0.02 0.43 ± 0.02 V_(max)  120 ± 3   79.5 ± 3.3  amylopectin K_(m) 0.26 ± 0.04 0.074 ± 0.008 V_(max) 84.5 ± 2.4  67.9 ± 1.7 

[0144] TABLE 14 Comparison of kinetic data for expressed SS's. Kinetic parameter SSI-3^(a) SSI-1 SSIIa-2 SSIIa-1 SSIIb-2^(a) SSIIb-1 K_(m) for amylopectin 0.24 0.15 0.13 0.15 0.26 0.07 K_(m) for glycogen — — 0.077 0.35 0.36 0.43 K_(m) for ADPGlc 0.33 0.18 0.28 0.83 0.32 0.40 V_(max) (with amylopectin) 26.3 22.5 15.4 41.1 84.5 67.9 V_(max) (with glycogen) 43.4 39.0 11.3 31.6 120 79.5 # could not be reached for this enzyme.

[0145] TABLE 15 The starch synthase activities of the chimerical enzymes. Generation of chimerical enzymes of maize starch synthase: the recombination of N-terminal extensions with C-terminal catalytic domains of starch synthase WX2 STSIIa STSIIb C-catalytic C-catalytic C-catalytic domain domain domain STSI N1-WX2 N1-C2 N1-C3 N-extension (+) (−) (+) (−) (+) (−) NRA  9.0  6.6 39.7  89.2 NRA STSIIa N2-WX2 N2-C2 N2-C3 N-extension (+) (−) (+) (−) (+) (−) 9.2 11.2 213.8  8.7 232.5 NRA STSIIb N3-WX2 N3-C2 N3-C3 N-extension (+) (−) (+) (−) (+) (−) NRA NRA  11.2 NRA 400.5 12.0

[0146] The photographs listed in the FIGS. 42 and 21 attempt to show the visual differences that are present into the starches as compared to those known in the art.

[0147] Description of the Starch

[0148] Corn starch is a milky, slightly thickened gel which is slightly if at all flowable.

[0149] Rice starch forms two levels the upper level is a thickened syrup like consistency more flowable then corn starch (less thick then corn starch) opaque milky color (more translucent then corn starch in this level) and a lower level which is a very white glob not transmitting much light through this bottom level of material. This lower level is formed in a very thick mass and does not appear flowable.

[0150] Corn amylopectin is slightly less white then the top level of rice starch and is a very slightly opaque milky color (more translucent then corn starch) slightly less flowable then the rice top level.

[0151] Potato dextrin is the most transparent almost appearing clear but is still opaque white and it is very flowable appearing only slightly less flowable then water.

[0152] Waxy Maize starch will flow very slowly and has the consistency of honey. The color is very opaque transmitting little light and the color is only slightly less light then corn starch.

[0153] SSI starch made from plasmid pExs-8 has two distinct levels. The top level appears clear and slightly thicker then the flowability of water. The bottom level appears as a precipitate. This sample resembles the ornaments that contain little figures; and plastic flakes resembling snowflakes. Like those ornaments when turned upside down the sample appears to be falling snow. However the flakes in this sample appear to be slightly gummy and appear in the first moments of level mixing to form a opaque white liquid.

[0154] SSI starch made from a host containing the following two plasmids pExsC BEI BEII and pExs8 is not as clear as the top level of pExs-8 and appears slightly less thick then pExs-8. It has even more flowability then does Potato Dextrin.

[0155] SSIIb starch made from a host containing the following two plasmids pExsC BEI BEII and pExs-9 is not as clear as the top level of pExs-8 and appears slightly less thick then pExs-8. It has even more flowability then does Potato Dextrin.

[0156] WAXY starch made from a host containing the following two plasmids pExsC BEI BEII and pExs-wx is not as clear as the top level of pExs-8 but seems to have a few tiny thread like chains that settle to the bottom and when mixed give the material a slightly, more white color and appears slightly less thick then pExs-8. It has even more flowability then does Potato Dextrin.

[0157] SSII starch made from a host containing the following two plasmids pExsC BEI BEII and pExs-3a is the color of corn starch and maybe slightly whiter but not as white as the bottom level of pExs8 and definitely transmitting more light through and has the flowability characteristic of pExs-8 when mixed.

[0158] glgA starch appears to have a very slight precipitate and is comparable in color to corn amylose pectin and ExsC BEI BEII and pExs-wx. And the flowability is between corn amylose and pExsC BEI BEII pExs-wx.

[0159] The samples of polysaccharides listed above form groups generally according to color as follows. waxy maize starch and cornstarch and pExsC BEI BEII pExs3a and pExsc8 are the whitest group. The flowability characteristics of this group are fairly diverse. With corn starch a lump and Waxy maize starch only slightly flowable and pExsC BEI BEII and pExs-3a and pExsC-8 more like water then syrup. The second group contains corn amylopectin and pExsC BEI BEII pExs-wx and pExsC BEI BEII and pExs-Al which are less white and clearer. The flowability of corn amylopectin is less then the other two members of this group but it is still similar. The last group is the least white and thus the clearest. This group includes pExsC BEI BEII and pExs-8, potato dextrin, pExsC BEI BEII and pExs-IO, pExsC BEI BEII and pExs-9. The flowability of this group is also similar to each other.

[0160] Plant Hosts

[0161] The following plasmids have been transformed into rice plants. The sequence for the mutant glgC gene is shown in FIG. 46. The plasmids are made substantially in a similar manner as described above for the production of bacterial plasmid. Clearly the plasmid maps shown in FIGS. 25-41 and this application and the listed short protocols allow the ordinarily skilled person in the art to make the present plasmids. The following combinations of plasmids have been transformed into rice plants. Additionally combinations of plasmids including the combination that includes all of the maize genes SSI, SSII, SSII, BEI, BEII, and GBSS in one host or alternatively in two host that are then crossed to form a hybrid having the entire complement of up regulated starch genes are being developed. Clearly the ordinarily skilled person in the art could have placed the sequences in the antisense positions to down regulate these genes to the extent that maize genes will down regulate the partial homologous ricer genes. The first group of transgenic are group, including rice transformants (transformed by microprojectile bombardment) containing MSTSI-2 (pExs52) and glgC₃ (pExs66), MSTSIIa-2 and glgC₃ (pExs53 and pExs56). The second group of rice transformants contains MSTSIIa and glgC₃ (pExs54 and pExs56). The third group of transformation contain: transgenic 5 MSTSIIb and glgC₃ (pExs 61 and pExs 66): transgenic 6 Maize wx and glgC₃ pExs65 and pExs66). Additionally, glgA and glgB and glgC are combined and transformed into rice. This last transformant is combining the rice plants starch pathway with the gene encoding for ADPG pyrophosphorylase and the bacterial genes. The combination of the plasmids encoding for at least one of the following enzymes, SSI, SSIIa, SSIIb, SSIII, Debranching enzymes, BEI, BEII, GBSS (wx) and some or all of the bacterial starch genes is also useful. There ae presently over 300 transformants in the greenhouse. The T1 transgenic rice plants have been screened and characterized (FIG. 56, 57). 12 plants have successfully expressed maize SSI-2 in rice seeds. 21 plants have successfully expressed maize SSIIb in rice seeds. We are currently screening rice plants down regulated the rice SS expression by cosuppression and have 400 T2 plants in the greenhouse.

[0162] Maize Starch Synthase and its Mutant Forms.

[0163] In order to characterize the multiple forms of maize starch synthase. the genes coding for the full length SS and its N-terminally truncated forms were expressed in E. coli. The recombinant enzymes were purified and kinetically characterized. We have demonstrated that different isoforms and its truncated forms all have distinct properties (Table 6-14, FIG. 58-63). The specific activities (Vmax) of the purified maize SSI-1, SSI-2, and SSI-3 were 22.5, 33.4 and 26.3:mol glc/min/mg of protein respectively, Our results have clearly indicated that the catalytic center of SSI is not located in its N-terminal extension. However. N-terminal truncation decreased the enzyme affinity for amylopectin, with the K_(m) for amylopectin of the truncated SSI-3 being about 60%-90% higher than that of the full length SSI-1. The effects of N-terminal truncation of SSIIa depend upon the assay conditions used. For both SSIIa-1 and SSIIa-2, the V_(max) of each enzyme increased 2-fold upon raising assay temperature from 27^(N)C to 37^(N)C (Tables II and III). However, the effect of temperature on ADPGlc affinity was different for SSIIa-1 and SSIIa-2. For the truncated SSIIa-2, the K_(m) for ADPGlc was not affected by raising temperature. In contrast, the K_(m) of ADPGlc for the putative full length SSIIa-I increased 2 fold upon raising the assay temperature from 27 NC to 37 NC (Table III). Interestingly, the truncated SSIIa-2 exhibited a lower K_(m) for ADPGlc than SSIIa-1 did in all assay conditions used in this study except that they showed similar K_(m) values for ADPGlc when glycogen was used as a primer at 27EC. Although N-terminal truncation of SSIIa appears to lower the K_(m) for ADPGlc under most assay conditions, it also must be noted that the maximal velocity of the truncated SSIIa-2 is decreased by about 2-4 fold when compared to SSIIa-1. The truncated SSIIb-2 was found, to be more temperature stable than the longer SSIIb-1 in the presence of citrate, while little difference was observed in their pH activity profiles. While the putative full length SSIIb-1 showed a similar V_(max) using amylopectin or glycogen as a primer, the N-terminally truncated SSIIb-2 showed a 40% increase in V_(max) using glycogen compared with amylopectin as a primer. N-terminal truncation of SSIIb increased its V_(max) by 25% with amylopectin as a primer. We also demonstrated that kimeric enzymes of maize starch synthase (combining the C-terminal domain of SS with different N-terminal sequences of SS or unrelated sequences would produce a functional enzyme with SS activity and altered properties) (Table 15).

CONCLUSIONS, RAMIFICATIONS, AND SCOPE

[0164] Accordingly, it can be seen that, according to the invention, The starch genes can produce new and altered starch in either host, plant or bacteria. Additionally, polysaccahrides very similar to corn starch can be produced in a bacterial host.

[0165] Although the description above contains many specificities, these should not be construed as limiting the scope of the invention but as merely providing illustrations of some of the presently preferred embodiments of this invention. Various other embodiments and ramifications are possible within it's scope. For example, different combinations of the plasmids in either host for the production of useful plant and useful grain and useful polysaccahrides.

[0166] Thus the scope of the invention should be determined by the appended claims and their legal equivalents, rather than by the examples given. All references cited herein are incorporated herein in their entirety by reference.

1 77 1 10 PRT Zea mays 1 Gly Glu Asn Val Met Asn Val Ile Val Val 1 5 10 2 9 PRT Zea mays 2 Ala Glu Ala Glu Ala Gly Gly Lys Asp 1 5 3 10 PRT Zea mays 3 Met Ser Ile Val Phe Val Thr Gly Glu Ala 1 5 10 4 9 PRT Zea mays 4 Gly Asp Leu Gly Leu Glu Pro Glu Gly 1 5 5 9 PRT Zea mays 5 Cys Val Ala Glu Leu Ser Arg Glu Gly 1 5 6 10 PRT Zea mays 6 Gly Ser Val Gly Ala Ala Leu Arg Ser Tyr 1 5 10 7 10 PRT Zea mays 7 Met Asn Val Val Val Val Ala Ser Glu Cys 1 5 10 8 10 PRT Zea mays 8 Ala Ser Ala Gly Met Asn Val Val Phe Val 1 5 10 9 10 PRT Zea mays 9 Met Asn Val Val Phe Val Gly Ala Glu Met 1 5 10 10 8 PRT Zea mays 10 Ala Ala Ala Pro Ala Gly Glu Glu 1 5 11 11 PRT Zea mays 11 Cys Val Ala Glu Leu Ser Arg Glu Gly Pro Ala 1 5 10 12 12 PRT Zea mays 12 Met Asn Val Val Val Val Ala Ser Glu Cys Ala Pro 1 5 10 13 15 PRT Zea mays 13 Ala Ser Ala Gly Met Asn Val Val Phe Val Gly Ala Glu Met Ala 1 5 10 15 14 11 PRT Zea mays 14 Met Asn Val Val Phe Val Gly Ala Glu Met Ala 1 5 10 15 13 PRT Zea mays 15 Met Ser Ile Val Phe Thr Gly Glu Ala Ser Pro Tyr Ala 1 5 10 16 15 PRT Zea mays 16 Cys Val Ala Glu Leu Ser Arg Asp Leu Gly Leu Glu Pro Glu Gly 1 5 10 15 17 5 PRT Zea mays MOD_RES (2) Ser or Thr 17 Lys Xaa Gly Gly Leu 1 5 18 19 DNA Artificial Sequence Description of Artificial Sequence primer 18 caagaatgct gcgggagtc 19 19 33 DNA Artificial Sequence Description of Artificial Sequence primer 19 aagtcgacat atgtgcgtcg cggagctgag cag 33 20 32 DNA Artificial Sequence Description of Artificial Sequence primer 20 gggccccata tgagcattgt ctttgtaacc gg 32 21 31 DNA Artificial Sequence Description of Artificial Sequence primer 21 ctcgggccca tatgggggag aatgttatga a 31 22 24 DNA Artificial Sequence Description of Artificial Sequence primer 22 gaggcatcaa tgaacacaaa gtcg 24 23 39 DNA Artificial Sequence Description of Artificial Sequence primer 23 gaagggcccc atatggctga ggctgaggcc gggggcaag 39 24 33 DNA Artificial Sequence Description of Artificial Sequence primer 24 ttggatccat atgggagctg cggttgcatt ggg 33 25 21 DNA Artificial Sequence Description of Artificial Sequence primer 25 cctgcgggct ctggcttcac c 21 26 33 DNA Artificial Sequence Description of Artificial Sequence primer 26 ttggatccat atgaacgtcg tcgtggtggc ttc 33 27 23 DNA Artificial Sequence Description of Artificial Sequence primer 27 gcataccatg gaacctcaac agc 23 28 27 DNA Artificial Sequence Description of Artificial Sequence primer 28 ggtaccatat gaacgtcgtc ttcggcg 27 29 22 DNA Artificial Sequence Description of Artificial Sequence primer 29 gacaggcccg tagatcttct cc 22 30 35 DNA Artificial Sequence Description of Artificial Sequence primer 30 ttggtaccat atggccagcg ccgccggcat gaacg 35 31 25 DNA Artificial Sequence Description of Artificial Sequence primer 31 gaagatctgg cagggacctg cacac 25 32 28 DNA Artificial Sequence Description of Artificial Sequence primer 32 ggactagtgc attatcgctc ctgtttat 28 33 1488 DNA E. coli CDS (join(1..1431, 1435..1488)) glgA 33 atg cag gtt tta cat gta tgt tca gag atg ttc ccg ctg ctt aaa acc 48 Met Gln Val Leu His Val Cys Ser Glu Met Phe Pro Leu Leu Lys Thr 1 5 10 15 ggc ggt ctg gct gat gtt att ggg gca tta ccc gca gca caa atc gca 96 Gly Gly Leu Ala Asp Val Ile Gly Ala Leu Pro Ala Ala Gln Ile Ala 20 25 30 gac ggc gtt gac gct cgc gta ctg ttg cct gca ttt ccc gac att cgc 144 Asp Gly Val Asp Ala Arg Val Leu Leu Pro Ala Phe Pro Asp Ile Arg 35 40 45 cgt ggc gtg acc gat gcg cag gta gta tcc cgt cgt gat acc tcc gcc 192 Arg Gly Val Thr Asp Ala Gln Val Val Ser Arg Arg Asp Thr Ser Ala 50 55 60 gga cat atc acg ctg ttg ttc ggt cat tac aac ggg gtt ggc att tac 240 Gly His Ile Thr Leu Leu Phe Gly His Tyr Asn Gly Val Gly Ile Tyr 65 70 75 80 ctg att gac gcg ccg cat ctc tat gat cgt ccg gga agt ccg tat cac 288 Leu Ile Asp Ala Pro His Leu Tyr Asp Arg Pro Gly Ser Pro Tyr His 85 90 95 gat acc aac tta ttt gtc cat acc gac aac gta ttg cgt ttt gcg ctg 336 Asp Thr Asn Leu Phe Val His Thr Asp Asn Val Leu Arg Phe Ala Leu 100 105 110 ctg ggg tgg gtt ggg gca gaa atg gcc agc ggg ctt gac cca ttc tgg 384 Leu Gly Trp Val Gly Ala Glu Met Ala Ser Gly Leu Asp Pro Phe Trp 115 120 125 cgt cct gat gtg gtg cat gcg cac gac tgg cat gca ggc ctt gcg cct 432 Arg Pro Asp Val Val His Ala His Asp Trp His Ala Gly Leu Ala Pro 130 135 140 gcg tat ctg gcg gcg cgc ggg cgt ccg gcg aag tcg gtg ttt act gtg 480 Ala Tyr Leu Ala Ala Arg Gly Arg Pro Ala Lys Ser Val Phe Thr Val 145 150 155 160 cac aac cta gcc tat caa ggc atg ttt tat gca cat cac atg aat gac 528 His Asn Leu Ala Tyr Gln Gly Met Phe Tyr Ala His His Met Asn Asp 165 170 175 atc caa ttg cca tgg tca ttc ttt aat att cat ggg ctg gaa ttc aac 576 Ile Gln Leu Pro Trp Ser Phe Phe Asn Ile His Gly Leu Glu Phe Asn 180 185 190 gga caa atc tct ttc ctg aag gcc ggt ctg tac tat gcc gat cac att 624 Gly Gln Ile Ser Phe Leu Lys Ala Gly Leu Tyr Tyr Ala Asp His Ile 195 200 205 acg gcg gtc agt cca acc tac gct cgc gag atc acc gaa ccg cag ttt 672 Thr Ala Val Ser Pro Thr Tyr Ala Arg Glu Ile Thr Glu Pro Gln Phe 210 215 220 gcc tac ggt atg gaa ggt ctg ttg caa cag cgt cac cgc gaa ggg cgt 720 Ala Tyr Gly Met Glu Gly Leu Leu Gln Gln Arg His Arg Glu Gly Arg 225 230 235 240 ctt tcc ggc gta ccg aac ggc gtg gac gag aaa atc tgg agt cca gag 768 Leu Ser Gly Val Pro Asn Gly Val Asp Glu Lys Ile Trp Ser Pro Glu 245 250 255 acg gac tta ctg ttg gcc tcg cgt tac acc cgc gat acg ttg gaa gat 816 Thr Asp Leu Leu Leu Ala Ser Arg Tyr Thr Arg Asp Thr Leu Glu Asp 260 265 270 aaa gcg gaa aat aag cgc cag tca caa atc gca atg gga tcc aag gtt 864 Lys Ala Glu Asn Lys Arg Gln Ser Gln Ile Ala Met Gly Ser Lys Val 275 280 285 gac gat aaa gtg ccg ctt ttt gca gtg gtg agc cgt ctg acc agc cag 912 Asp Asp Lys Val Pro Leu Phe Ala Val Val Ser Arg Leu Thr Ser Gln 290 295 300 aaa ggt ctc gat tcg gtg ctg gaa gcc tca ccg ggt tct tcg gag cag 960 Lys Gly Leu Asp Ser Val Leu Glu Ala Ser Pro Gly Ser Ser Glu Gln 305 310 315 320 ggc ggg cag ctg gcg cta ctc ggc gcg ggc gat ccg gtg ctg cag gaa 1008 Gly Gly Gln Leu Ala Leu Leu Gly Ala Gly Asp Pro Val Leu Gln Glu 325 330 335 ggt ttc ctt gcg gcg gca gcg gaa tac ccc ggt cag gtg ggc gtt cag 1056 Gly Phe Leu Ala Ala Ala Ala Glu Tyr Pro Gly Gln Val Gly Val Gln 340 345 350 att ggc tat cac gaa gca ttt tcg cat cgc att atg ggc ggc gcg gac 1104 Ile Gly Tyr His Glu Ala Phe Ser His Arg Ile Met Gly Gly Ala Asp 355 360 365 gtc att ctg gtg ccc agc cgt ttc gaa ccg tgc ggc tta acg caa ctt 1152 Val Ile Leu Val Pro Ser Arg Phe Glu Pro Cys Gly Leu Thr Gln Leu 370 375 380 tat gga tcg aag tac ggt acg ctg ccg tta gtg cga cgc acc ggt ggg 1200 Tyr Gly Ser Lys Tyr Gly Thr Leu Pro Leu Val Arg Arg Thr Gl Gly 385 390 395 400 ctt gct gat acg gtt tct gac tgt tct ctc gag aac ctt gca gat ggc 1248 Leu Ala Asp Thr Val Ser Asp Cys Ser Leu Glu Asn Leu Ala Asp Gly 405 410 415 gtc gcc aat ggg ttt atc ttc gaa gat agt aat gcc tgg tcg ctg tta 1296 Val Ala Asn Gly Phe Ile Phe Glu Asp Ser Asn Ala Trp Ser Leu Leu 420 425 430 cgg act att cga cgt gct ttt gta ctg tgg tcc tgt cct cca ctg tgg 1344 Arg Thr Ile Arg Arg Ala Phe Val Leu Trp Ser Cys Pro Pro Leu Trp 435 440 445 cgg ttt gtg caa cgt cag gct atg gca atg gat ttt ggc tgg cag gtc 1392 Arg Phe Val Gln Arg Gln Ala Met Ala Met Asp Phe Gly Trp Gln Val 450 455 460 gcg gcg aag tcg tac cgt gag ctt tac tat cgc tcg aaa tag ttt tca 1440 Ala Ala Lys Ser Tyr Arg Glu Leu Tyr Tyr Arg Ser Lys Phe Ser 465 470 475 gga aac gcc tac atg aat gct ccg ttt aca tat tca tcg ccc acg ctt 1488 Gly Asn Ala Tyr Met Asn Ala Pro Phe Thr Tyr Ser Ser Pro Thr Leu 480 485 490 495 34 495 PRT E. coli glgA 34 Met Gln Val Leu His Val Cys Ser Glu Met Phe Pro Leu Leu Lys Thr 1 5 10 15 Gly Gly Leu Ala Asp Val Ile Gly Ala Leu Pro Ala Ala Gln Ile Ala 20 25 30 Asp Gly Val Asp Ala Arg Val Leu Leu Pro Ala Phe Pro Asp Ile Arg 35 40 45 Arg Gly Val Thr Asp Ala Gln Val Val Ser Arg Arg Asp Thr Ser Ala 50 55 60 Gly His Ile Thr Leu Leu Phe Gly His Tyr Asn Gly Val Gly Ile Tyr 65 70 75 80 Leu Ile Asp Ala Pro His Leu Tyr Asp Arg Pro Gly Ser Pro Tyr His 85 90 95 Asp Thr Asn Leu Phe Val His Thr Asp Asn Val Leu Arg Phe Ala Leu 100 105 110 Leu Gly Trp Val Gly Ala Glu Met Ala Ser Gly Leu Asp Pro Phe Trp 115 120 125 Arg Pro Asp Val Val His Ala His Asp Trp His Ala Gly Leu Ala Pro 130 135 140 Ala Tyr Leu Ala Ala Arg Gly Arg Pro Ala Lys Ser Val Phe Thr Val 145 150 155 160 His Asn Leu Ala Tyr Gln Gly Met Phe Tyr Ala His His Met Asn Asp 165 170 175 Ile Gln Leu Pro Trp Ser Phe Phe Asn Ile His Gly Leu Glu Phe Asn 180 185 190 Gly Gln Ile Ser Phe Leu Lys Ala Gly Leu Tyr Tyr Ala Asp His Ile 195 200 205 Thr Ala Val Ser Pro Thr Tyr Ala Arg Glu Ile Thr Glu Pro Gln Phe 210 215 220 Ala Tyr Gly Met Glu Gly Leu Leu Gln Gln Arg His Arg Glu Gly Arg 225 230 235 240 Leu Ser Gly Val Pro Asn Gly Val Asp Glu Lys Ile Trp Ser Pro Glu 245 250 255 Thr Asp Leu Leu Leu Ala Ser Arg Tyr Thr Arg Asp Thr Leu Glu Asp 260 265 270 Lys Ala Glu Asn Lys Arg Gln Ser Gln Ile Ala Met Gly Ser Lys Val 275 280 285 Asp Asp Lys Val Pro Leu Phe Ala Val Val Ser Arg Leu Thr Ser Gln 290 295 300 Lys Gly Leu Asp Ser Val Leu Glu Ala Ser Pro Gly Ser Ser Glu Gln 305 310 315 320 Gly Gly Gln Leu Ala Leu Leu Gly Ala Gly Asp Pro Val Leu Gln Glu 325 330 335 Gly Phe Leu Ala Ala Ala Ala Glu Tyr Pro Gly Gln Val Gly Val Gln 340 345 350 Ile Gly Tyr His Glu Ala Phe Ser His Arg Ile Met Gly Gly Ala Asp 355 360 365 Val Ile Leu Val Pro Ser Arg Phe Glu Pro Cys Gly Leu Thr Gln Leu 370 375 380 Tyr Gly Ser Lys Tyr Gly Thr Leu Pro Leu Val Arg Arg Thr Gly Gly 385 390 395 400 Leu Ala Asp Thr Val Ser Asp Cys Ser Leu Glu Asn Leu Ala Asp Gly 405 410 415 Val Ala Asn Gly Phe Ile Phe Glu Asp Ser Asn Ala Trp Ser Leu Leu 420 425 430 Arg Thr Ile Arg Arg Ala Phe Val Leu Trp Ser Cys Pro Pro Leu Trp 435 440 445 Arg Phe Val Gln Arg Gln Ala Met Ala Met Asp Phe Gly Trp Gln Val 450 455 460 Ala Ala Lys Ser Tyr Arg Glu Leu Tyr Tyr Arg Ser Lys Phe Ser Gly 465 470 475 480 Asn Ala Tyr Met Asn Ala Pro Phe Thr Tyr Ser Ser Pro Thr Leu 485 490 495 35 2361 DNA E. coli glgB 35 atg tcc gat cgt atc gat aga gac gtg att aac gcg cta att gca ggc 48 Met Ser Asp Arg Ile Asp Arg Asp Val Ile Asn Ala Leu Ile Ala Gly 1 5 10 15 cat ttt gcg gat cct ttt tcc gta ctg gga atg cat aaa acc acc gcg 96 His Phe Ala Asp Pro Phe Ser Val Leu Gly Met His Lys Thr Thr Ala 20 25 30 gga ctg gaa gtc cgt gcc ctt tta ccc gac gct acc gat gtg tgg gtg 144 Gly Leu Glu Val Arg Ala Leu Leu Pro Asp Ala Thr Asp Val Trp Val 35 40 45 att gaa ccg aaa acc ggg cgc aaa ctc gca aaa ctg gag tgt ctc gac 192 Ile Glu Pro Lys Thr Gly Arg Lys Leu Ala Lys Leu Glu Cys Leu Asp 50 55 60 tca cgg gga ttc ttt agc ggc gtc att ccg cga cgt aag aat ttt ttc 240 Ser Arg Gly Phe Phe Ser Gly Val Ile Pro Arg Arg Lys Asn Phe Phe 65 70 75 80 cgc tat cag ttg gct gtt gtc tgg cat ggt cag caa aac ctg att gat 288 Arg Tyr Gln Leu Ala Val Val Trp His Gly Gln Gln Asn Leu Ile Asp 85 90 95 gat cct tac cgt ttt ggt ccg cta atc cag gaa atg gat gcc tgg cta 336 Asp Pro Tyr Arg Phe Gly Pro Leu Ile Gln Glu Met Asp Ala Trp Leu 100 105 110 tta tct gaa ggt act cac ctg cgc ccg tat gaa acc tta ggc gcg cat 384 Leu Ser Glu Gly Thr His Leu Arg Pro Tyr Glu Thr Leu Gly Ala His 115 120 125 gca gat act atg gat ggc gtc aca ggt acg cgt ttc tct gtc tgg gct 432 Ala Asp Thr Met Asp Gly Val Thr Gly Thr Arg Phe Ser Val Trp Ala 130 135 140 cca aac gcc cgt cgg gtc tcg gtg gtt ggg caa ttc aac tac tgg gac 480 Pro Asn Ala Arg Arg Val Ser Val Val Gly Gln Phe Asn Tyr Trp Asp 145 150 155 160 ggt cgc cgt cac ccg atg cgc ctg cgt aaa gag agc ggc atc tgg gaa 528 Gly Arg Arg His Pro Met Arg Leu Arg Lys Glu Ser Gly Ile Trp Glu 165 170 175 ctg ttt atc cct ggg gcg cat aac ggt cag ctc tat aaa tac gag atg 576 Leu Phe Ile Pro Gly Ala His Asn Gly Gln Leu Tyr Lys Tyr Glu Met 180 185 190 att gat gcc aat ggc aac ttg cgt ctg aag tcc gac cct tat gcc ttt 624 Ile Asp Ala Asn Gly Asn Leu Arg Leu Lys Ser Asp Pro Tyr Ala Phe 195 200 205 gaa gcg caa atg cgc ccg gaa acc gcg tct ctt att tgc ggg ctg ccg 672 Glu Ala Gln Met Arg Pro Glu Thr Ala Ser Leu Ile Cys Gly Leu Pro 210 215 220 gaa aag gtt gta cag act gaa gag cgc aaa aaa gcg aat cag ttt gat 720 Glu Lys Val Val Gln Thr Glu Glu Arg Lys Lys Ala Asn Gln Phe Asp 225 230 235 240 gcg cca atc tct att tat gaa gtt cac ctg ggt tcc tgg cgt cgc cac 768 Ala Pro Ile Ser Ile Tyr Glu Val His Leu Gly Ser Trp Arg Arg His 245 250 255 acc gac aac aat ttc tgg ttg agc tac cgc gag ctg gcc gat caa ctg 816 Thr Asp Asn Asn Phe Trp Leu Ser Tyr Arg Glu Leu Ala Asp Gln Leu 260 265 270 gtg cct tat gct aaa tgg atg ggc ttt acc cac ctc gaa cta ctg ccc 864 Val Pro Tyr Ala Lys Trp Met Gly Phe Thr His Leu Glu Leu Leu Pro 275 280 285 att aac gag cat ccc ttc gat ggc agt tgg ggt tat cag cca acc ggc 912 Ile Asn Glu His Pro Phe Asp Gly Ser Trp Gly Tyr Gln Pro Thr Gly 290 295 300 ctg tat gcg cca acc cgc cgt ttt ggt act cgc gac gac ttc cgt tat 960 Leu Tyr Ala Pro Thr Arg Arg Phe Gly Thr Arg Asp Asp Phe Arg Tyr 305 310 315 320 ttc att gat gcc gca cac gca gct ggt ctg aac gtg att ctc gac tgg 1008 Phe Ile Asp Ala Ala His Ala Ala Gly Leu Asn Val Ile Leu Asp Trp 325 330 335 gtg cca ggc cac ttc ccg act gat gac ttt gcg ctt gcc gaa ttt gat 1056 Val Pro Gly His Phe Pro Thr Asp Asp Phe Ala Leu Ala Glu Phe Asp 340 345 350 ggc acg aac ttg tat gaa cac agc gat ccg cgt gaa ggc tat cat cag 1104 Gly Thr Asn Leu Tyr Glu His Ser Asp Pro Arg Glu Gly Tyr His Gln 355 360 365 gac tgg aac acg ctg atc tac aac tat ggt cgc cgt gaa gtc agt aac 1152 Asp Trp Asn Thr Leu Ile Tyr Asn Tyr Gly Arg Arg Glu Val Ser Asn 370 375 380 ttc ctc gtc ggt aac gcg ctt tac tgg att gaa cgt ttt ggt att gat 1200 Phe Leu Val Gly Asn Ala Leu Tyr Trp Ile Glu Arg Phe Gly Ile Asp 385 390 395 400 gcg ctg cgc gtc gat gcg gtg gcg tca atg att tat cgc gac tac agc 1248 Ala Leu Arg Val Asp Ala Val Ala Ser Met Ile Tyr Arg Asp Tyr Ser 405 410 415 cgt aaa gag ggg gag tgg atc ccg aac gaa ttt ggc ggg cgc gag aat 1296 Arg Lys Glu Gly Glu Trp Ile Pro Asn Glu Phe Gly Gly Arg Glu Asn 420 425 430 ctt gaa gcg att gaa ttc ttg cgt aat acc aac cgt att ctt ggt gag 1344 Leu Glu Ala Ile Glu Phe Leu Arg Asn Thr Asn Arg Ile Leu Gly Glu 435 440 445 cag gtt tcc ggt gcg gtg aca atg gct gag gag tct acc gat ttc cct 1392 Gln Val Ser Gly Ala Val Thr Met Ala Glu Glu Ser Thr Asp Phe Pro 450 455 460 ggc gtt tct cgt ccg cag gat atg ggc ggt ctg ggc ttc tgg tac aag 1440 Gly Val Ser Arg Pro Gln Asp Met Gly Gly Leu Gly Phe Trp Tyr Lys 465 470 475 480 tgg aac ctc ggc tgg atg cat gac acc ttg gac tac atg aag ctc gac 1488 Trp Asn Leu Gly Trp Met His Asp Thr Leu Asp Tyr Met Lys Leu Asp 485 490 495 ccg gtt tat cgt cag tat cat cac gat aaa ctg acc ttc ggg att ctc 1536 Pro Val Tyr Arg Gln Tyr His His Asp Lys Leu Thr Phe Gly Ile Leu 500 505 510 tac aac tac act gaa aac ttc gtc ctg ccg ttg tcg cat gat gaa gtg 1584 Tyr Asn Tyr Thr Glu Asn Phe Val Leu Pro Leu Ser His Asp Glu Val 515 520 525 gtc cac ggt aaa aaa tcg att ctc gac cgc atg ccg ggc gac gca tgg 1632 Val His Gly Lys Lys Ser Ile Leu Asp Arg Met Pro Gly Asp Ala Trp 530 535 540 cag aaa ttc gcg aac ctg cgc gcc tac tat ggc tgg atg tgg gca ttc 1680 Gln Lys Phe Ala Asn Leu Arg Ala Tyr Tyr Gly Trp Met Trp Ala Phe 545 550 555 560 ccg ggc aag aaa cta ctg ttc atg ggt aac gaa ttt gcc cag ggc cgc 1728 Pro Gly Lys Lys Leu Leu Phe Met Gly Asn Glu Phe Ala Gln Gly Arg 565 570 575 gag tgg aac cat gac gcc agc ctc gac tgg cat ctg ttg gaa ggc ggc 1776 Glu Trp Asn His Asp Ala Ser Leu Asp Trp His Leu Leu Glu Gly Gly 580 585 590 gat aac tgg cac cac ggt gtc cag cgt ctg gtg cgc gat ctg aac ctc 1824 Asp Asn Trp His His Gly Val Gln Arg Leu Val Arg Asp Leu Asn Leu 595 600 605 acc tac cgc cac cat aaa gca atg cat gaa ctg gat ttt gac ccg tac 1872 Thr Tyr Arg His His Lys Ala Met His Glu Leu Asp Phe Asp Pro Tyr 610 615 620 ggc ttt gaa tgg ctg gtg gtg gat gac aaa gaa cgc tcg gtg ctg atc 1920 Gly Phe Glu Trp Leu Val Val Asp Asp Lys Glu Arg Ser Val Leu Ile 625 630 635 640 ttt gtg cgt cgc gat aaa gag ggt aac gaa atc atc gtt gcc agt aac 1968 Phe Val Arg Arg Asp Lys Glu Gly Asn Glu Ile Ile Val Ala Ser Asn 645 650 655 ttt acg ccg gta ccg cgt cat gat tat cgc ttc ggc ata aac cag ccg 2016 Phe Thr Pro Val Pro Arg His Asp Tyr Arg Phe Gly Ile Asn Gln Pro 660 665 670 ggc aaa tgg cgt gaa atc ctc aat acc gat tcc atg cac tat cac ggc 2064 Gly Lys Trp Arg Glu Ile Leu Asn Thr Asp Ser Met His Tyr His Gly 675 680 685 agt aat gca ggc aat ggc ggc acg gta cac agc gat gag att gcc agc 2112 Ser Asn Ala Gly Asn Gly Gly Thr Val His Ser Asp Glu Ile Ala Ser 690 695 700 cac ggt cgt cag cat tca cta agc ctg acg cta cca ccg ctg gcc act 2160 His Gly Arg Gln His Ser Leu Ser Leu Thr Leu Pro Pro Leu Ala Thr 705 710 715 720 atc tgg ctg gtt cgg gag gca gaa tga cac aac tcg cca ttg gca aac 2208 Ile Trp Leu Val Arg Glu Ala Glu His Asn Ser Pro Leu Ala Asn 725 730 735 ccg ctc ccc tcg gcg cgc att acg acg gtc agg gcg tca act tca cac 2256 Pro Leu Pro Ser Ala Arg Ile Thr Thr Val Arg Ala Ser Thr Ser His 740 745 750 ttt tct ccg ctc atg ccg agc ggg tag aac tgt gtg tct ttg acg cca 2304 Phe Ser Pro Leu Met Pro Ser Gly Asn Cys Val Ser Leu Thr Pro 755 760 765 atg gcc agg aac atc gct atg act tgc cag ggc aca gtg gcg aca ttt 2352 Met Ala Arg Asn Ile Ala Met Thr Cys Gln Gly Thr Val Ala Thr Phe 770 775 780 ggc acg gtt 2361 Gly Thr Val 785 36 785 PRT E. coli glgB 36 Met Ser Asp Arg Ile Asp Arg Asp Val Ile Asn Ala Leu Ile Ala Gly 1 5 10 15 His Phe Ala Asp Pro Phe Ser Val Leu Gly Met His Lys Thr Thr Ala 20 25 30 Gly Leu Glu Val Arg Ala Leu Leu Pro Asp Ala Thr Asp Val Trp Val 35 40 45 Ile Glu Pro Lys Thr Gly Arg Lys Leu Ala Lys Leu Glu Cys Leu Asp 50 55 60 Ser Arg Gly Phe Phe Ser Gly Val Ile Pro Arg Arg Lys Asn Phe Phe 65 70 75 80 Arg Tyr Gln Leu Ala Val Val Trp His Gly Gln Gln Asn Leu Ile Asp 85 90 95 Asp Pro Tyr Arg Phe Gly Pro Leu Ile Gln Glu Met Asp Ala Trp Leu 100 105 110 Leu Ser Glu Gly Thr His Leu Arg Pro Tyr Glu Thr Leu Gly Ala His 115 120 125 Ala Asp Thr Met Asp Gly Val Thr Gly Thr Arg Phe Ser Val Trp Ala 130 135 140 Pro Asn Ala Arg Arg Val Ser Val Val Gly Gln Phe Asn Tyr Trp Asp 145 150 155 160 Gly Arg Arg His Pro Met Arg Leu Arg Lys Glu Ser Gly Ile Trp Glu 165 170 175 Leu Phe Ile Pro Gly Ala His Asn Gly Gln Leu Tyr Lys Tyr Glu Met 180 185 190 Ile Asp Ala Asn Gly Asn Leu Arg Leu Lys Ser Asp Pro Tyr Ala Phe 195 200 205 Glu Ala Gln Met Arg Pro Glu Thr Ala Ser Leu Ile Cys Gly Leu Pro 210 215 220 Glu Lys Val Val Gln Thr Glu Glu Arg Lys Lys Ala Asn Gln Phe Asp 225 230 235 240 Ala Pro Ile Ser Ile Tyr Glu Val His Leu Gly Ser Trp Arg Arg His 245 250 255 Thr Asp Asn Asn Phe Trp Leu Ser Tyr Arg Glu Leu Ala Asp Gln Leu 260 265 270 Val Pro Tyr Ala Lys Trp Met Gly Phe Thr His Leu Glu Leu Leu Pro 275 280 285 Ile Asn Glu His Pro Phe Asp Gly Ser Trp Gly Tyr Gln Pro Thr Gly 290 295 300 Leu Tyr Ala Pro Thr Arg Arg Phe Gly Thr Arg Asp Asp Phe Arg Tyr 305 310 315 320 Phe Ile Asp Ala Ala His Ala Ala Gly Leu Asn Val Ile Leu Asp Trp 325 330 335 Val Pro Gly His Phe Pro Thr Asp Asp Phe Ala Leu Ala Glu Phe Asp 340 345 350 Gly Thr Asn Leu Tyr Glu His Ser Asp Pro Arg Glu Gly Tyr His Gln 355 360 365 Asp Trp Asn Thr Leu Ile Tyr Asn Tyr Gly Arg Arg Glu Val Ser Asn 370 375 380 Phe Leu Val Gly Asn Ala Leu Tyr Trp Ile Glu Arg Phe Gly Ile Asp 385 390 395 400 Ala Leu Arg Val Asp Ala Val Ala Ser Met Ile Tyr Arg Asp Tyr Ser 405 410 415 Arg Lys Glu Gly Glu Trp Ile Pro Asn Glu Phe Gly Gly Arg Glu Asn 420 425 430 Leu Glu Ala Ile Glu Phe Leu Arg Asn Thr Asn Arg Ile Leu Gly Glu 435 440 445 Gln Val Ser Gly Ala Val Thr Met Ala Glu Glu Ser Thr Asp Phe Pro 450 455 460 Gly Val Ser Arg Pro Gln Asp Met Gly Gly Leu Gly Phe Trp Tyr Lys 465 470 475 480 Trp Asn Leu Gly Trp Met His Asp Thr Leu Asp Tyr Met Lys Leu Asp 485 490 495 Pro Val Tyr Arg Gln Tyr His His Asp Lys Leu Thr Phe Gly Ile Leu 500 505 510 Tyr Asn Tyr Thr Glu Asn Phe Val Leu Pro Leu Ser His Asp Glu Val 515 520 525 Val His Gly Lys Lys Ser Ile Leu Asp Arg Met Pro Gly Asp Ala Trp 530 535 540 Gln Lys Phe Ala Asn Leu Arg Ala Tyr Tyr Gly Trp Met Trp Ala Phe 545 550 555 560 Pro Gly Lys Lys Leu Leu Phe Met Gly Asn Glu Phe Ala Gln Gly Arg 565 570 575 Glu Trp Asn His Asp Ala Ser Leu Asp Trp His Leu Leu Glu Gly Gly 580 585 590 Asp Asn Trp His His Gly Val Gln Arg Leu Val Arg Asp Leu Asn Leu 595 600 605 Thr Tyr Arg His His Lys Ala Met His Glu Leu Asp Phe Asp Pro Tyr 610 615 620 Gly Phe Glu Trp Leu Val Val Asp Asp Lys Glu Arg Ser Val Leu Ile 625 630 635 640 Phe Val Arg Arg Asp Lys Glu Gly Asn Glu Ile Ile Val Ala Ser Asn 645 650 655 Phe Thr Pro Val Pro Arg His Asp Tyr Arg Phe Gly Ile Asn Gln Pro 660 665 670 Gly Lys Trp Arg Glu Ile Leu Asn Thr Asp Ser Met His Tyr His Gly 675 680 685 Ser Asn Ala Gly Asn Gly Gly Thr Val His Ser Asp Glu Ile Ala Ser 690 695 700 His Gly Arg Gln His Ser Leu Ser Leu Thr Leu Pro Pro Leu Ala Thr 705 710 715 720 Ile Trp Leu Val Arg Glu Ala Glu His Asn Ser Pro Leu Ala Asn Pro 725 730 735 Leu Pro Ser Ala Arg Ile Thr Thr Val Arg Ala Ser Thr Ser His Phe 740 745 750 Ser Pro Leu Met Pro Ser Gly Asn Cys Val Ser Leu Thr Pro Met Ala 755 760 765 Arg Asn Ile Ala Met Thr Cys Gln Gly Thr Val Ala Thr Phe Gly Thr 770 775 780 Val 785 37 150 PRT Zea mays 37 Met Ala Ala Lys Met Leu Ala Leu Phe Ala Leu Leu Ala Leu Cys Ala 1 5 10 15 Ser Ala Thr Ser Ala Thr His Ile Pro Gly His Leu Pro Pro Val Met 20 25 30 Pro Leu Gly Thr Met Asn Pro Cys Met Gln Tyr Cys Met Met Gln Gln 35 40 45 Gly Leu Ala Ser Leu Met Ala Cys Pro Ser Leu Met Leu Gln Gln Leu 50 55 60 Leu Ala Leu Pro Leu Gln Thr Met Pro Val Met Met Pro Gln Met Met 65 70 75 80 Thr Pro Asn Met Met Ser Pro Leu Met Met Pro Ser Met Met Ser Pro 85 90 95 Met Val Leu Pro Ser Met Met Ser Gln Ile Met Met Pro Gln Cys His 100 105 110 Cys Asp Ala Val Ser Gln Ile Met Leu Gln Gln Gln Leu Pro Phe Met 115 120 125 Phe Asn Pro Met Ala Met Thr Ile Pro Pro Met Phe Leu Gln Gln Pro 130 135 140 Phe Val Gly Ala Ala Phe 145 150 38 2562 DNA Zea mays 38 aagcttgcta ctttctttcc ttaatgttga tttccccttt gttagatgtt ctttgtgtta 60 tatacactct gtatacaagg atgcgataca cacatcagct agtcctaatg atgccaccga 120 ctttacttga ggaaaaggaa acaaatatga tgtggccatc acattctcaa taacaatgac 180 catgtgcgca atgacatacc atcatatttg atatcataaa aataaattta ttatcaaagt 240 aaacatatag ttcatatatc agatattaaa gtgataagaa caaatattac attttatctt 300 atataaaatg acgaaaaagg tacgagttga aaaggagtcc aacccctttt ttatagcttg 360 ttcggttgct tgttctcttc ggctagcgag gtggtagaat gtgagagtgt tgcgcgtgga 420 ttcccgtcgt agtgttctta ggtgatttct cacggcccat ctgtgatata gcgactcata 480 tgtggtgtaa tagcccattg ggagaagggg agagatatag atctacgtga tttgcacgtg 540 atgcacgacg aacgaaactg gtggtttaaa gtagtagagg tttgtcatta gaggtgtaaa 600 tggtacatat attatccgtt catattcgaa tttgatccgt ataagagggc taagatctaa 660 tccgtataca agtccaagta ttaagtatcc gatccatatc ggatctttat ccgtatccgt 720 atactcaaaa tttgatgttt aagattttaa tatatattta aactttatag gaactcgata 780 atatttgtat ctgatttgaa ttatgaaaac aaatatggaa cgattaattt cagtctatat 840 ccgttccgat atttgtcatg ctttgctaaa aataccttta caaggcatct tgtgcagatt 900 atatattaat ctgaaatcag ttagagaagc ctacaaattt gaccaaatgc cgagtcatcc 960 ggcttatccc ctttccaact ttcagttctg caagcgccag aaatcgtttt tcatctacat 1020 tgtctttgtt gcctgcatac atctataaat aggacctgct agatcaatcg cagtccatcg 1080 gcctcagtcg cacatatcta ctatactata ctctaggaag caaggacacc accgccatgg 1140 cagccaagat gcttgcattg ttcgctctcc tagctctttg tgcaagcgcc actagtgcga 1200 cccatattcc agggcacttg ccaccagtca tgccattggg taccatgaac ccatgcatgc 1260 agtactgcat gatgcaacag gggcttgcca gcttgatggc gtgtccgtcc ctgatgctgc 1320 agcaactgtt ggccttaccg cttcagacga tgccagtgat gatgccacag atgatgacgc 1380 ctaacatgat gtcaccattg atgatgccga gcatgatgtc accaatggtc ttgccgagca 1440 tgatgtcgca aataatgatg ccacaatgtc actgcgacgc cgtctcgcag attatgctgc 1500 aacagcagtt accattcatg ttcaacccaa tggccatgac gattccaccc atgttcttac 1560 agcaaccctt tgttggtgct gcattctaga tagaaatatt tgtgttgtat cgaataatga 1620 gttgacatgc catcgcgtgt gactcattat taacaataaa acaagtttcc tcttattatc 1680 tttttatatc tctccctatc catttttgca aagcccatta tcctttactc cctaagtccc 1740 aatatatttt agaccttaaa ttgtatgtct atattcaaaa gaatgacaat aaatctagac 1800 atatatataa aacacataca ttaagtattg tatgaatcta ttaaaatgct aaaacgacta 1860 atattatggg acggagggag tactttatta gtagattaca ttgttatttt ctctattcca 1920 aatataagtc tggtttttca atcaatcaat atatattacc atgtccaaac attttgaatt 1980 atatatctag gtgcagcatc cgtgcacgat cgtaaaagaa gcagtcacgg tgttggtccc 2040 aaaaactaat cgtccgttgt cggtcaccta taaagattca tgaagagaac caaaataagg 2100 caatataatt aatgtaatat gactcctcct tttgaattac ttaggaataa cataagcaaa 2160 caaaaaaagg agaagatcaa ggtaaataaa ggcattttgt gagaaaacat ggaagcataa 2220 gaatgcataa gtaatgattt gtgtctcttt atattttttt tattcacgtg aatttacata 2280 gataccatcg gatgttcgat ggtaatacaa tgatgcctta gctccgagag cttcgaatga 2340 tgagcgattt aaaaatactc ctatcaattg ttcgaaagtt ctttgtctca tgcatgggca 2400 atgtacctct atttataggg acggtgcgac gtacaaattt gtataaaatt atatttttat 2460 tcccaaatcc tatgcatatg tgtcggggac cataattagg ggtaccctca aggctcctaa 2520 ttctcagctg gtaaccccat cagcataaag ctgcaaaggc ct 2562 39 1141 DNA Zea mays 39 aagcttgcta ctttctttcc ttaatgttga tttccccttt gttagatgtt ctttgtgtta 60 tatacactct gtatacaagg atgcgataca cacatcagct agtcctaatg atgccaccga 120 ctttacttga ggaaaaggaa acaaatatga tgtggccatc acattctcaa taacaatgac 180 catgtgcgca atgacatacc atcatatttg atatcataaa aataaattta ttatcaaagt 240 aaacatatag ttcatatatc agatattaaa gtgataagaa caaatattac attttatctt 300 atataaaatg acgaaaaagg tacgagttga aaaggagtcc aacccctttt ttatagcttg 360 ttcggttgct tgttctcttc ggctagcgag gtggtagaat gtgagagtgt tgcgcgtgga 420 ttcccgtcgt agtgttctta ggtgatttct cacggcccat ctgtgatata gcgactcata 480 tatgtggtgt aatagcccat tgggagaagg ggagagatat agatctacgt gatttgcacg 540 tgatgcacga cgaacgaaac tggtggttta aagtagtaga ggtttgtcat tagaggtgta 600 aatggtacat atattatccg ttcatattcg aatttgatcc gtataagagg gctaagatct 660 aatccgtata caagtccaag tattaagtat ccgatccata tcggatcttt atccgtatcc 720 gtatactcaa aatttgatgt ttaagatttt aatatatatt taaactttat aggaactcga 780 taatatttgt atctgatttg aattatgaaa acaaatatgg aacgattaat ttcagtctat 840 atccgttccg atatttgtca tgctttgcta aaaatacctt tacaaggcat cttgtgcaga 900 ttatatatta atctgaaatc agttagagaa gcctacaaat ttgaccaaat gccgagtcat 960 ccggcttatc ccctttccaa ctttcagttc tgcaagcgcc agaaatcgtt tttcatctac 1020 attgtctttg ttgcctgcat acatctataa ataggacctg ctagatcaat cgcagtccat 1080 cggcctcagt cgcacatatc tactatacta tactctagga agcaaggaca ccaccgccat 1140 g 1141 40 1328 DNA E. coli glgC3 40 atg gtt agt tta gag aag aac gat cac tta atg ttg gcg cgc cag ctg 48 Met Val Ser Leu Glu Lys Asn Asp His Leu Met Leu Ala Arg Gln Leu 1 5 10 15 cca ttg aaa tct gtt gcc ctg ata ctg gcg gga gga cgt ggt acc cgc 96 Pro Leu Lys Ser Val Ala Leu Ile Leu Ala Gly Gly Arg Gly Thr Arg 20 25 30 ctg aag gat tta acc aat aag cga gca aaa ccg gcc gta cac ttc ggc 144 Leu Lys Asp Leu Thr Asn Lys Arg Ala Lys Pro Ala Val His Phe Gly 35 40 45 ggt aag ttc cgc att atc gac ttt gcg ctg tct aac tgc atc aac tcc 192 Gly Lys Phe Arg Ile Ile Asp Phe Ala Leu Ser Asn Cys Ile Asn Ser 50 55 60 ggg atc cgt cgt atg ggc gtg atc acc cag tac cag tcc cac act ctg 240 Gly Ile Arg Arg Met Gly Val Ile Thr Gln Tyr Gln Ser His Thr Leu 65 70 75 80 gtg cag cac att cag cgc ggc tgg tca ttc ttc aat gaa gaa atg aac 288 Val Gln His Ile Gln Arg Gly Trp Ser Phe Phe Asn Glu Glu Met Asn 85 90 95 gag ttt gtc gat ctg ctg cca gca cag cag aga atg aaa ggg gaa aac 336 Glu Phe Val Asp Leu Leu Pro Ala Gln Gln Arg Met Lys Gly Glu Asn 100 105 110 tgg tat cgc ggc acc gca gat gcg gtc acc caa aac ctc gac att atc 384 Trp Tyr Arg Gly Thr Ala Asp Ala Val Thr Gln Asn Leu Asp Ile Ile 115 120 125 cgt cgt tat aaa gcg gaa tac gtg gtg atc ctg gcg ggc gac cat atc 432 Arg Arg Tyr Lys Ala Glu Tyr Val Val Ile Leu Ala Gly Asp His Ile 130 135 140 tac aag caa gac tac tcg cgt atg ctt atc gat cac gtc gaa aaa ggt 480 Tyr Lys Gln Asp Tyr Ser Arg Met Leu Ile Asp His Val Glu Lys Gly 145 150 155 160 gta cgt tgt acc gtt gtt tgt atg cca gta ccg att gaa gaa gcc tcc 528 Val Arg Cys Thr Val Val Cys Met Pro Val Pro Ile Glu Glu Ala Ser 165 170 175 gca ttt ggc gtt atg gcg gtt gat gag aac gat aaa act atc gaa ttc 576 Ala Phe Gly Val Met Ala Val Asp Glu Asn Asp Lys Thr Ile Glu Phe 180 185 190 gtg gaa aaa cct gct aac ccg ccg tca atg ccg aac gat ccg agc aaa 624 Val Glu Lys Pro Ala Asn Pro Pro Ser Met Pro Asn Asp Pro Ser Lys 195 200 205 tct ctg gcg agt atg ggt atc tac gtc ttt gac gcc gac tat ctg tat 672 Ser Leu Ala Ser Met Gly Ile Tyr Val Phe Asp Ala Asp Tyr Leu Tyr 210 215 220 gaa ctg ctg gaa gaa gac gat cgc gat gag aac tcc agc cac gac ttt 720 Glu Leu Leu Glu Glu Asp Asp Arg Asp Glu Asn Ser Ser His Asp Phe 225 230 235 240 ggc aaa gat ttg att ccc aag atc acc gaa gcc ggt ctg gcc tat gcg 768 Gly Lys Asp Leu Ile Pro Lys Ile Thr Glu Ala Gly Leu Ala Tyr Ala 245 250 255 cac ccg ttc ccg ctc tct tgc gta caa tcc gac ccg gat gcc gag ccg 816 His Pro Phe Pro Leu Ser Cys Val Gln Ser Asp Pro Asp Ala Glu Pro 260 265 270 tac tgg cgc gat gtg ggt acg ctg gaa gct tac tgg aaa gcg aac ctc 864 Tyr Trp Arg Asp Val Gly Thr Leu Glu Ala Tyr Trp Lys Ala Asn Leu 275 280 285 gat ctg gcc tct gtg gtg gac aaa ctg gat atg tac gat cgc aat tgg 912 Asp Leu Ala Ser Val Val Asp Lys Leu Asp Met Tyr Asp Arg Asn Trp 290 295 300 cca att cgc acc tac aat gaa tca tta ccg cca gcg aaa ttc gtg cag 960 Pro Ile Arg Thr Tyr Asn Glu Ser Leu Pro Pro Ala Lys Phe Val Gln 305 310 315 320 gat cgc tcc ggt agc cac ggg atg acc ctt aac tca ctg gtt tcc ggc 1008 Asp Arg Ser Gly Ser His Gly Met Thr Leu Asn Ser Leu Val Ser Gly 325 330 335 ggt tgt gtg atc tcc ggt tcg gtg gtg gtg cag tcc gtt ctg ttc tcg 1056 Gly Cys Val Ile Ser Gly Ser Val Val Val Gln Ser Val Leu Phe Ser 340 345 350 cgc gtt cgc gtg aat tca ttc tgc aac att gat tcc gcc gta ttg tta 1104 Arg Val Arg Val Asn Ser Phe Cys Asn Ile Asp Ser Ala Val Leu Leu 355 360 365 ccg gaa gta tgg gta ggt cgc tcg tgc cgt ctg cgc cgc tgc gtc atc 1152 Pro Glu Val Trp Val Gly Arg Ser Cys Arg Leu Arg Arg Cys Val Ile 370 375 380 gat cgt gct tgt gtt att ccg gaa ggc atg gtg att ggt gaa aac gca 1200 Asp Arg Ala Cys Val Ile Pro Glu Gly Met Val Ile Gly Glu Asn Ala 385 390 395 400 gag gaa gat gca cgt cgt ttc tat cgt tca gaa gaa ggc atc gtg ctg 1248 Glu Glu Asp Ala Arg Arg Phe Tyr Arg Ser Glu Glu Gly Ile Val Leu 405 410 415 gta acg cgc gaa atg cta cgg aag tta ggg cat aaa cag gag cga 1293 Val Thr Arg Glu Met Leu Arg Lys Leu Gly His Lys Gln Glu Arg 420 425 430 taa tgc agg ttt tac atg tat gtt cag aga tgt tt 1328 Cys Arg Phe Tyr Met Tyr Val Gln Arg Cys 435 440 41 441 PRT E. coli glgC3 41 Met Val Ser Leu Glu Lys Asn Asp His Leu Met Leu Ala Arg Gln Leu 1 5 10 15 Pro Leu Lys Ser Val Ala Leu Ile Leu Ala Gly Gly Arg Gly Thr Arg 20 25 30 Leu Lys Asp Leu Thr Asn Lys Arg Ala Lys Pro Ala Val His Phe Gly 35 40 45 Gly Lys Phe Arg Ile Ile Asp Phe Ala Leu Ser Asn Cys Ile Asn Ser 50 55 60 Gly Ile Arg Arg Met Gly Val Ile Thr Gln Tyr Gln Ser His Thr Leu 65 70 75 80 Val Gln His Ile Gln Arg Gly Trp Ser Phe Phe Asn Glu Glu Met Asn 85 90 95 Glu Phe Val Asp Leu Leu Pro Ala Gln Gln Arg Met Lys Gly Glu Asn 100 105 110 Trp Tyr Arg Gly Thr Ala Asp Ala Val Thr Gln Asn Leu Asp Ile Ile 115 120 125 Arg Arg Tyr Lys Ala Glu Tyr Val Val Ile Leu Ala Gly Asp His Ile 130 135 140 Tyr Lys Gln Asp Tyr Ser Arg Met Leu Ile Asp His Val Glu Lys Gly 145 150 155 160 Val Arg Cys Thr Val Val Cys Met Pro Val Pro Ile Glu Glu Ala Ser 165 170 175 Ala Phe Gly Val Met Ala Val Asp Glu Asn Asp Lys Thr Ile Glu Phe 180 185 190 Val Glu Lys Pro Ala Asn Pro Pro Ser Met Pro Asn Asp Pro Ser Lys 195 200 205 Ser Leu Ala Ser Met Gly Ile Tyr Val Phe Asp Ala Asp Tyr Leu Tyr 210 215 220 Glu Leu Leu Glu Glu Asp Asp Arg Asp Glu Asn Ser Ser His Asp Phe 225 230 235 240 Gly Lys Asp Leu Ile Pro Lys Ile Thr Glu Ala Gly Leu Ala Tyr Ala 245 250 255 His Pro Phe Pro Leu Ser Cys Val Gln Ser Asp Pro Asp Ala Glu Pro 260 265 270 Tyr Trp Arg Asp Val Gly Thr Leu Glu Ala Tyr Trp Lys Ala Asn Leu 275 280 285 Asp Leu Ala Ser Val Val Asp Lys Leu Asp Met Tyr Asp Arg Asn Trp 290 295 300 Pro Ile Arg Thr Tyr Asn Glu Ser Leu Pro Pro Ala Lys Phe Val Gln 305 310 315 320 Asp Arg Ser Gly Ser His Gly Met Thr Leu Asn Ser Leu Val Ser Gly 325 330 335 Gly Cys Val Ile Ser Gly Ser Val Val Val Gln Ser Val Leu Phe Ser 340 345 350 Arg Val Arg Val Asn Ser Phe Cys Asn Ile Asp Ser Ala Val Leu Leu 355 360 365 Pro Glu Val Trp Val Gly Arg Ser Cys Arg Leu Arg Arg Cys Val Ile 370 375 380 Asp Arg Ala Cys Val Ile Pro Glu Gly Met Val Ile Gly Glu Asn Ala 385 390 395 400 Glu Glu Asp Ala Arg Arg Phe Tyr Arg Ser Glu Glu Gly Ile Val Leu 405 410 415 Val Thr Arg Glu Met Leu Arg Lys Leu Gly His Lys Gln Glu Arg Cys 420 425 430 Arg Phe Tyr Met Tyr Val Gln Arg Cys 435 440 42 1328 DNA E. coli glcC 42 atg gtt agt tta gag aag aac gat cac tta atg ttg gcg cgc cag ctg 48 Met Val Ser Leu Glu Lys Asn Asp His Leu Met Leu Ala Arg Gln Leu 1 5 10 15 cca ttg aaa tct gtt gcc ctg ata ctg gcg gga gga cgt ggt acc cgc 96 Pro Leu Lys Ser Val Ala Leu Ile Leu Ala Gly Gly Arg Gly Thr Arg 20 25 30 ctg aag gat tta acc aat aag cga gca aaa ccg gcc gta cac ttc ggc 144 Leu Lys Asp Leu Thr Asn Lys Arg Ala Lys Pro Ala Val His Phe Gly 35 40 45 ggt aag ttc cgc att atc gac ttt gcg ctg tct aac tgc atc aac tcc 192 Gly Lys Phe Arg Ile Ile Asp Phe Ala Leu Ser Asn Cys Ile Asn Ser 50 55 60 ggg atc cgt cgt atg ggc gtg atc acc cag tac cag tcc cac act ctg 240 Gly Ile Arg Arg Met Gly Val Ile Thr Gln Tyr Gln Ser His Thr Leu 65 70 75 80 gtg cag cac att cag cgc ggc tgg tca ttc ttc aat gaa gaa atg aac 288 Val Gln His Ile Gln Arg Gly Trp Ser Phe Phe Asn Glu Glu Met Asn 85 90 95 gag ttt gtc gat ctg ctg cca gca cag cag aga atg aaa ggg gaa aac 336 Glu Phe Val Asp Leu Leu Pro Ala Gln Gln Arg Met Lys Gly Glu Asn 100 105 110 tgg tat cgc ggc acc gca gat gcg gtc acc caa aac ctc gac att atc 384 Trp Tyr Arg Gly Thr Ala Asp Ala Val Thr Gln Asn Leu Asp Ile Ile 115 120 125 cgc cgt tat aaa gcg gaa tac gtg gtg atc ctg gcg ggc gac cat atc 432 Arg Arg Tyr Lys Ala Glu Tyr Val Val Ile Leu Ala Gly Asp His Ile 130 135 140 tac aag caa gac tac tcg cgt atg ctt atc gat cac gtc gaa aaa ggc 480 Tyr Lys Gln Asp Tyr Ser Arg Met Leu Ile Asp His Val Glu Lys Gly 145 150 155 160 gca cgt tgc acc gtt gct tgt atg cca gta ccg att gaa gaa gcc tcc 528 Ala Arg Cys Thr Val Ala Cys Met Pro Val Pro Ile Glu Glu Ala Ser 165 170 175 gca ttt ggc gtt atg gcg gtt gat gag aac gat aaa att atc gaa ttc 576 Ala Phe Gly Val Met Ala Val Asp Glu Asn Asp Lys Ile Ile Glu Phe 180 185 190 gtt gaa aaa cct gct aac ccg ccg tca atg ccg aac gat ccg agc aaa 624 Val Glu Lys Pro Ala Asn Pro Pro Ser Met Pro Asn Asp Pro Ser Lys 195 200 205 tct ctg gcg agt atg ggt atc tac gtc ttt gac gcc gac tat ctg tat 672 Ser Leu Ala Ser Met Gly Ile Tyr Val Phe Asp Ala Asp Tyr Leu Tyr 210 215 220 gaa ctg ctg gaa gaa gac gat cgc gat gag aac tcc agc cac gac ttt 720 Glu Leu Leu Glu Glu Asp Asp Arg Asp Glu Asn Ser Ser His Asp Phe 225 230 235 240 ggc aaa gat ttg att ccc aag atc acc gaa gcc ggt ctg gcc tat gcg 768 Gly Lys Asp Leu Ile Pro Lys Ile Thr Glu Ala Gly Leu Ala Tyr Ala 245 250 255 cac ccg ttc ccg ctc tct tgc gta caa tcc gac ccg gat gcc gag ccg 816 His Pro Phe Pro Leu Ser Cys Val Gln Ser Asp Pro Asp Ala Glu Pro 260 265 270 tac tgg cgc gat gtg ggt acg ctg gaa gct tac tgg aaa gcg aac ctc 864 Tyr Trp Arg Asp Val Gly Thr Leu Glu Ala Tyr Trp Lys Ala Asn Leu 275 280 285 gat ctg gcc tct gtg gtg ccg gaa ctg gat atg tac gat cgc aat tgg 912 Asp Leu Ala Ser Val Val Pro Glu Leu Asp Met Tyr Asp Arg Asn Trp 290 295 300 cca att cgc acc tac aat gaa tca tta ccg cca gcg aaa ttc gtg cag 960 Pro Ile Arg Thr Tyr Asn Glu Ser Leu Pro Pro Ala Lys Phe Val Gln 305 310 315 320 gat cgc tcc ggt agc cac ggg atg acc ctt aac tca ctg gtt tcc ggc 1008 Asp Arg Ser Gly Ser His Gly Met Thr Leu Asn Ser Leu Val Ser Gly 325 330 335 ggt tgt gtg atc tcc ggt tcg gtg gtg gtg cag tcc gtt ctg ttc tcg 1056 Gly Cys Val Ile Ser Gly Ser Val Val Val Gln Ser Val Leu Phe Ser 340 345 350 cgc gtt cgc gtg aat tca ttc tgc aac att gat tcc gcc gta ttg tta 1104 Arg Val Arg Val Asn Ser Phe Cys Asn Ile Asp Ser Ala Val Leu Leu 355 360 365 ccg gaa gta tgg gta ggt cgc tcg tgc cgt ctg cgc cgc tgc gtc atc 1152 Pro Glu Val Trp Val Gly Arg Ser Cys Arg Leu Arg Arg Cys Val Ile 370 375 380 gat cgt gct tgt gtt att ccg gaa ggc atg gtg att ggt gaa aac gca 1200 Asp Arg Ala Cys Val Ile Pro Glu Gly Met Val Ile Gly Glu Asn Ala 385 390 395 400 gag gaa gat gca cgt cgt ttc tat cgt tca gaa gaa ggc atc gtg ctg 1248 Glu Glu Asp Ala Arg Arg Phe Tyr Arg Ser Glu Glu Gly Ile Val Leu 405 410 415 gta acg cgc gaa atg cta cgg aag tta ggg cat aaa cag gag cga 1293 Val Thr Arg Glu Met Leu Arg Lys Leu Gly His Lys Gln Glu Arg 420 425 430 taa tgc agg ttt tac atg tat gtt cag aga tgt tt 1328 Cys Arg Phe Tyr Met Tyr Val Gln Arg Cys 435 440 43 441 PRT E. coli glcC 43 Met Val Ser Leu Glu Lys Asn Asp His Leu Met Leu Ala Arg Gln Leu 1 5 10 15 Pro Leu Lys Ser Val Ala Leu Ile Leu Ala Gly Gly Arg Gly Thr Arg 20 25 30 Leu Lys Asp Leu Thr Asn Lys Arg Ala Lys Pro Ala Val His Phe Gly 35 40 45 Gly Lys Phe Arg Ile Ile Asp Phe Ala Leu Ser Asn Cys Ile Asn Ser 50 55 60 Gly Ile Arg Arg Met Gly Val Ile Thr Gln Tyr Gln Ser His Thr Leu 65 70 75 80 Val Gln His Ile Gln Arg Gly Trp Ser Phe Phe Asn Glu Glu Met Asn 85 90 95 Glu Phe Val Asp Leu Leu Pro Ala Gln Gln Arg Met Lys Gly Glu Asn 100 105 110 Trp Tyr Arg Gly Thr Ala Asp Ala Val Thr Gln Asn Leu Asp Ile Ile 115 120 125 Arg Arg Tyr Lys Ala Glu Tyr Val Val Ile Leu Ala Gly Asp His Ile 130 135 140 Tyr Lys Gln Asp Tyr Ser Arg Met Leu Ile Asp His Val Glu Lys Gly 145 150 155 160 Ala Arg Cys Thr Val Ala Cys Met Pro Val Pro Ile Glu Glu Ala Ser 165 170 175 Ala Phe Gly Val Met Ala Val Asp Glu Asn Asp Lys Ile Ile Glu Phe 180 185 190 Val Glu Lys Pro Ala Asn Pro Pro Ser Met Pro Asn Asp Pro Ser Lys 195 200 205 Ser Leu Ala Ser Met Gly Ile Tyr Val Phe Asp Ala Asp Tyr Leu Tyr 210 215 220 Glu Leu Leu Glu Glu Asp Asp Arg Asp Glu Asn Ser Ser His Asp Phe 225 230 235 240 Gly Lys Asp Leu Ile Pro Lys Ile Thr Glu Ala Gly Leu Ala Tyr Ala 245 250 255 His Pro Phe Pro Leu Ser Cys Val Gln Ser Asp Pro Asp Ala Glu Pro 260 265 270 Tyr Trp Arg Asp Val Gly Thr Leu Glu Ala Tyr Trp Lys Ala Asn Leu 275 280 285 Asp Leu Ala Ser Val Val Pro Glu Leu Asp Met Tyr Asp Arg Asn Trp 290 295 300 Pro Ile Arg Thr Tyr Asn Glu Ser Leu Pro Pro Ala Lys Phe Val Gln 305 310 315 320 Asp Arg Ser Gly Ser His Gly Met Thr Leu Asn Ser Leu Val Ser Gly 325 330 335 Gly Cys Val Ile Ser Gly Ser Val Val Val Gln Ser Val Leu Phe Ser 340 345 350 Arg Val Arg Val Asn Ser Phe Cys Asn Ile Asp Ser Ala Val Leu Leu 355 360 365 Pro Glu Val Trp Val Gly Arg Ser Cys Arg Leu Arg Arg Cys Val Ile 370 375 380 Asp Arg Ala Cys Val Ile Pro Glu Gly Met Val Ile Gly Glu Asn Ala 385 390 395 400 Glu Glu Asp Ala Arg Arg Phe Tyr Arg Ser Glu Glu Gly Ile Val Leu 405 410 415 Val Thr Arg Glu Met Leu Arg Lys Leu Gly His Lys Gln Glu Arg Cys 420 425 430 Arg Phe Tyr Met Tyr Val Gln Arg Cys 435 440 44 1328 DNA E. coli glgCwt 44 atg gtt agt tta gag aag aac gat cac tta atg ttg gcg cgc cag ctg 48 Met Val Ser Leu Glu Lys Asn Asp His Leu Met Leu Ala Arg Gln Leu 1 5 10 15 cca ttg aaa tct gtt gcc ctg ata ctg gcg gga gga cgt ggt acc cgc 96 Pro Leu Lys Ser Val Ala Leu Ile Leu Ala Gly Gly Arg Gly Thr Arg 20 25 30 ctg aag gat tta acc aat aag cga gca aaa ccg gcc gta cac ttc ggc 144 Leu Lys Asp Leu Thr Asn Lys Arg Ala Lys Pro Ala Val His Phe Gly 35 40 45 ggt aag ttc cgc att atc gac ttt gcg ctg tct aac tgc atc aac tcc 192 Gly Lys Phe Arg Ile Ile Asp Phe Ala Leu Ser Asn Cys Ile Asn Ser 50 55 60 ggg atc cgt cgt atg ggc gtg atc acc cag tac cag tcc cac act ctg 240 Gly Ile Arg Arg Met Gly Val Ile Thr Gln Tyr Gln Ser His Thr Leu 65 70 75 80 gtg cag cac att cag cgc ggc tgg tca ttc ttc aat gaa gaa atg aac 288 Val Gln His Ile Gln Arg Gly Trp Ser Phe Phe Asn Glu Glu Met Asn 85 90 95 gag ttt gtc gat ctg ctg cca gca cag cag aga atg aaa ggg gaa aac 336 Glu Phe Val Asp Leu Leu Pro Ala Gln Gln Arg Met Lys Gly Glu Asn 100 105 110 tgg tat cgc ggc acc gca gat gcg gtc acc caa aac ctc gac att atc 384 Trp Tyr Arg Gly Thr Ala Asp Ala Val Thr Gln Asn Leu Asp Ile Ile 115 120 125 cgc cgt tat aaa gcg gaa tac gtg gtg atc ctg gcg ggc gac cat atc 432 Arg Arg Tyr Lys Ala Glu Tyr Val Val Ile Leu Ala Gly Asp His Ile 130 135 140 tac aag caa gac tac tcg cgt atg ctt atc gat cac gtc gaa aaa ggc 480 Tyr Lys Gln Asp Tyr Ser Arg Met Leu Ile Asp His Val Glu Lys Gly 145 150 155 160 gca cgt tgc acc gtt gct tgt atg cca gta ccg att gaa gaa gcc tcc 528 Ala Arg Cys Thr Val Ala Cys Met Pro Val Pro Ile Glu Glu Ala Ser 165 170 175 gca ttt ggc gtt atg gcg gtt gat gag aac gat aaa att atc gaa ttc 576 Ala Phe Gly Val Met Ala Val Asp Glu Asn Asp Lys Ile Ile Glu Phe 180 185 190 gtt gaa aaa cct gct aac ccg ccg tca atg ccg aac gat ccg agc aaa 624 Val Glu Lys Pro Ala Asn Pro Pro Ser Met Pro Asn Asp Pro Ser Lys 195 200 205 tct ctg gcg agt atg ggt atc tac gtc ttt gac gcc gac tat ctg tat 672 Ser Leu Ala Ser Met Gly Ile Tyr Val Phe Asp Ala Asp Tyr Leu Tyr 210 215 220 gaa ctg ctg gaa gaa gac gat cgc gat gag aac tcc agc cac gac ttt 720 Glu Leu Leu Glu Glu Asp Asp Arg Asp Glu Asn Ser Ser His Asp Phe 225 230 235 240 ggc aaa gat ttg att ccc aag atc acc gaa gcc ggt ctg gcc tat gcg 768 Gly Lys Asp Leu Ile Pro Lys Ile Thr Glu Ala Gly Leu Ala Tyr Ala 245 250 255 cac ccg ttc ccg ctc tct tgc gta caa tcc gac ccg gat gcc gag ccg 816 His Pro Phe Pro Leu Ser Cys Val Gln Ser Asp Pro Asp Ala Glu Pro 260 265 270 tac tgg cgc gat gtg ggt acg ctg gaa gct tac tgg aaa gcg aac ctc 864 Tyr Trp Arg Asp Val Gly Thr Leu Glu Ala Tyr Trp Lys Ala Asn Leu 275 280 285 gat ctg gcc tct gtg gtg ccg aaa ctg gat atg tac gat cgc aat tgg 912 Asp Leu Ala Ser Val Val Pro Lys Leu Asp Met Tyr Asp Arg Asn Trp 290 295 300 cca att cgc acc tac aat gaa tca tta ccg cca gcg aaa ttc gtg cag 960 Pro Ile Arg Thr Tyr Asn Glu Ser Leu Pro Pro Ala Lys Phe Val Gln 305 310 315 320 gat cgc tcc ggt agc cac ggg atg acc ctt aac tca ctg gtt tcc ggc 1008 Asp Arg Ser Gly Ser His Gly Met Thr Leu Asn Ser Leu Val Ser Gly 325 330 335 ggt tgt gtg atc tcc ggt tcg gtg gtg gtg cag tcc gtt ctg ttc tcg 1056 Gly Cys Val Ile Ser Gly Ser Val Val Val Gln Ser Val Leu Phe Ser 340 345 350 cgc gtt cgc gtg aat tca ttc tgc aac att gat tcc gcc gta ttg tta 1104 Arg Val Arg Val Asn Ser Phe Cys Asn Ile Asp Ser Ala Val Leu Leu 355 360 365 ccg gaa gta tgg gta ggt cgc tcg tgc cgt ctg cgc cgc tgc gtc atc 1152 Pro Glu Val Trp Val Gly Arg Ser Cys Arg Leu Arg Arg Cys Val Ile 370 375 380 gat cgt gct tgt gtt att ccg gaa ggc atg gtg att ggt gaa aac gca 1200 Asp Arg Ala Cys Val Ile Pro Glu Gly Met Val Ile Gly Glu Asn Ala 385 390 395 400 gag gaa gat gca cgt cgt ttc tat cgt tca gaa gaa ggc atc gtg ctg 1248 Glu Glu Asp Ala Arg Arg Phe Tyr Arg Ser Glu Glu Gly Ile Val Leu 405 410 415 gta acg cgc gaa atg cta cgg aag tta ggg cat aaa cag gag cga 1293 Val Thr Arg Glu Met Leu Arg Lys Leu Gly His Lys Gln Glu Arg 420 425 430 taa tgc agg ttt tac atg tat gtt cag aga tgt tt 1328 Cys Arg Phe Tyr Met Tyr Val Gln Arg Cys 435 440 45 441 PRT E. coli glgCwt 45 Met Val Ser Leu Glu Lys Asn Asp His Leu Met Leu Ala Arg Gln Leu 1 5 10 15 Pro Leu Lys Ser Val Ala Leu Ile Leu Ala Gly Gly Arg Gly Thr Arg 20 25 30 Leu Lys Asp Leu Thr Asn Lys Arg Ala Lys Pro Ala Val His Phe Gly 35 40 45 Gly Lys Phe Arg Ile Ile Asp Phe Ala Leu Ser Asn Cys Ile Asn Ser 50 55 60 Gly Ile Arg Arg Met Gly Val Ile Thr Gln Tyr Gln Ser His Thr Leu 65 70 75 80 Val Gln His Ile Gln Arg Gly Trp Ser Phe Phe Asn Glu Glu Met Asn 85 90 95 Glu Phe Val Asp Leu Leu Pro Ala Gln Gln Arg Met Lys Gly Glu Asn 100 105 110 Trp Tyr Arg Gly Thr Ala Asp Ala Val Thr Gln Asn Leu Asp Ile Ile 115 120 125 Arg Arg Tyr Lys Ala Glu Tyr Val Val Ile Leu Ala Gly Asp His Ile 130 135 140 Tyr Lys Gln Asp Tyr Ser Arg Met Leu Ile Asp His Val Glu Lys Gly 145 150 155 160 Ala Arg Cys Thr Val Ala Cys Met Pro Val Pro Ile Glu Glu Ala Ser 165 170 175 Ala Phe Gly Val Met Ala Val Asp Glu Asn Asp Lys Ile Ile Glu Phe 180 185 190 Val Glu Lys Pro Ala Asn Pro Pro Ser Met Pro Asn Asp Pro Ser Lys 195 200 205 Ser Leu Ala Ser Met Gly Ile Tyr Val Phe Asp Ala Asp Tyr Leu Tyr 210 215 220 Glu Leu Leu Glu Glu Asp Asp Arg Asp Glu Asn Ser Ser His Asp Phe 225 230 235 240 Gly Lys Asp Leu Ile Pro Lys Ile Thr Glu Ala Gly Leu Ala Tyr Ala 245 250 255 His Pro Phe Pro Leu Ser Cys Val Gln Ser Asp Pro Asp Ala Glu Pro 260 265 270 Tyr Trp Arg Asp Val Gly Thr Leu Glu Ala Tyr Trp Lys Ala Asn Leu 275 280 285 Asp Leu Ala Ser Val Val Pro Lys Leu Asp Met Tyr Asp Arg Asn Trp 290 295 300 Pro Ile Arg Thr Tyr Asn Glu Ser Leu Pro Pro Ala Lys Phe Val Gln 305 310 315 320 Asp Arg Ser Gly Ser His Gly Met Thr Leu Asn Ser Leu Val Ser Gly 325 330 335 Gly Cys Val Ile Ser Gly Ser Val Val Val Gln Ser Val Leu Phe Ser 340 345 350 Arg Val Arg Val Asn Ser Phe Cys Asn Ile Asp Ser Ala Val Leu Leu 355 360 365 Pro Glu Val Trp Val Gly Arg Ser Cys Arg Leu Arg Arg Cys Val Ile 370 375 380 Asp Arg Ala Cys Val Ile Pro Glu Gly Met Val Ile Gly Glu Asn Ala 385 390 395 400 Glu Glu Asp Ala Arg Arg Phe Tyr Arg Ser Glu Glu Gly Ile Val Leu 405 410 415 Val Thr Arg Glu Met Leu Arg Lys Leu Gly His Lys Gln Glu Arg Cys 420 425 430 Arg Phe Tyr Met Tyr Val Gln Arg Cys 435 440 46 1915 DNA Zea mays CDS (join(1..1815, 1819..1914)) 46 atg gcg gct ctg gcc acg tcg cag ctc gtc gca acg cgc gcc ggc ctg 48 Met Ala Ala Leu Ala Thr Ser Gln Leu Val Ala Thr Arg Ala Gly Leu 1 5 10 15 ggc gtc ccg gac gcg tcc acg ttc cgc cgc ggc gcc gcg cag ggc ctg 96 Gly Val Pro Asp Ala Ser Thr Phe Arg Arg Gly Ala Ala Gln Gly Leu 20 25 30 agg ggg gcc cgg gcg tcg gcg gcg gcg gac acg ctc agc atg cgg acc 144 Arg Gly Ala Arg Ala Ser Ala Ala Ala Asp Thr Leu Ser Met Arg Thr 35 40 45 agc gcg cgc gcg gcg ccc agg cac cag cag cag gcg gcg gcg ccc ccg 192 Ser Ala Arg Ala Ala Pro Arg His Gln Gln Gln Ala Ala Ala Pro Pro 50 55 60 tcc aag ggc agc gag cag cac acg gcc agc gcc ggc atg aac gtc gtc 240 Ser Lys Gly Ser Glu Gln His Thr Ala Ser Ala Gly Met Asn Val Val 65 70 75 80 ttc gtc ggc gcc gag atg gcg ccg tgg agc aag acc ggc ggc ctc ggc 288 Phe Val Gly Ala Glu Met Ala Pro Trp Ser Lys Thr Gly Gly Leu Gly 85 90 95 gac gtc ctc ggc ggc ctg ccg ccg gcc atg gcc gcg aac ggg cac cgt 336 Asp Val Leu Gly Gly Leu Pro Pro Ala Met Ala Ala Asn Gly His Arg 100 105 110 gtc atg gtc gtc tct ccc cgc tac gac cag tac aag gac gcc tgg gac 384 Val Met Val Val Ser Pro Arg Tyr Asp Gln Tyr Lys Asp Ala Trp Asp 115 120 125 acc agc gtc gtg tcc gag atc aag atg gga gac ggg tac gag acg gtc 432 Thr Ser Val Val Ser Glu Ile Lys Met Gly Asp Gly Tyr Glu Thr Val 130 135 140 agg ttc ttc cac tgc tac aag cgc gga gtg gac cgc gtg ttc gtt gac 480 Arg Phe Phe His Cys Tyr Lys Arg Gly Val Asp Arg Val Phe Val Asp 145 150 155 160 cac cca ctg ttc ctg gag agg gtt tgg gga aag acc gag gag aag atc 528 His Pro Leu Phe Leu Glu Arg Val Trp Gly Lys Thr Glu Glu Lys Ile 165 170 175 tac ggg cct gtc gct gga acg gac tac agg gac aac cag ctg cgg ttc 576 Tyr Gly Pro Val Ala Gly Thr Asp Tyr Arg Asp Asn Gln Leu Arg Phe 180 185 190 agc ctg cta tgc cag gga gca ctt gaa gct cca agg atc ctg agc ctc 624 Ser Leu Leu Cys Gln Gly Ala Leu Glu Ala Pro Arg Ile Leu Ser Leu 195 200 205 aac aac aac cca tac ttc tcc gga cca tac ggg agg gac gtc gtg ttc 672 Asn Asn Asn Pro Tyr Phe Ser Gly Pro Tyr Gly Arg Asp Val Val Phe 210 215 220 gtc tgc aac gac tgg cac acc ggc cct ctc tcg tgc tac ctc aag agc 720 Val Cys Asn Asp Trp His Thr Gly Pro Leu Ser Cys Tyr Leu Lys Ser 225 230 235 240 aac tac cag tcc cac ggc atc tac agg gac gca aag acc gct ttc tgc 768 Asn Tyr Gln Ser His Gly Ile Tyr Arg Asp Ala Lys Thr Ala Phe Cys 245 250 255 atc cac aac atc tcc tac cag ggc cgg ttc gcc ttc tcc gac tac ccg 816 Ile His Asn Ile Ser Tyr Gln Gly Arg Phe Ala Phe Ser Asp Tyr Pro 260 265 270 gag ctg aac ctc ccg gag aga ttc aag tcg tcc ttc gat ttc atc gac 864 Glu Leu Asn Leu Pro Glu Arg Phe Lys Ser Ser Phe Asp Phe Ile Asp 275 280 285 ggc tac gag aag ccc gtg gaa ggc cgg aag atc aac tgg atg aag gcc 912 Gly Tyr Glu Lys Pro Val Glu Gly Arg Lys Ile Asn Trp Met Lys Ala 290 295 300 ggg atc ctc gag gcc gac agg gtc ctc acc gtc agc ccc tac tac gcc 960 Gly Ile Leu Glu Ala Asp Arg Val Leu Thr Val Ser Pro Tyr Tyr Ala 305 310 315 320 gag gag ctc atc tcc ggc atc gcc agg ggc tgc gag ctc gac aac atc 1008 Glu Glu Leu Ile Ser Gly Ile Ala Arg Gly Cys Glu Leu Asp Asn Ile 325 330 335 atg cgc ctc acc ggc atc acc ggc atc gtc aac ggc atg gac gtc agc 1056 Met Arg Leu Thr Gly Ile Thr Gly Ile Val Asn Gly Met Asp Val Ser 340 345 350 gag tgg gac ccc agc agg gac aag tac atc gcc gtg aag tac gac gtg 1104 Glu Trp Asp Pro Ser Arg Asp Lys Tyr Ile Ala Val Lys Tyr Asp Val 355 360 365 tcg acg gcc gtg gag gcc aag gcg ctg aac aag gag gcg ctg cag gcg 1152 Ser Thr Ala Val Glu Ala Lys Ala Leu Asn Lys Glu Ala Leu Gln Ala 370 375 380 gag gtc ggg ctc ccg gtg gac cgg aac atc ccg ctg gtg gcg ttc atc 1200 Glu Val Gly Leu Pro Val Asp Arg Asn Ile Pro Leu Val Ala Phe Ile 385 390 395 400 ggc agg ctg gaa gag cag aag ggc ccc gac gtc atg gcg gcc gcc atc 1248 Gly Arg Leu Glu Glu Gln Lys Gly Pro Asp Val Met Ala Ala Ala Ile 405 410 415 ccg cag ctc atg gag atg gtg gag gac gtg cag atc gtt ctg ctg ggc 1296 Pro Gln Leu Met Glu Met Val Glu Asp Val Gln Ile Val Leu Leu Gly 420 425 430 acg ggc aag aag aag ttc gag cgc atg ctc atg agc gcc gag gag aag 1344 Thr Gly Lys Lys Lys Phe Glu Arg Met Leu Met Ser Ala Glu Glu Lys 435 440 445 ttc cca ggc aag gtg cgc gcc gtg gtc aag ttc aac gcg gcg ctg gcg 1392 Phe Pro Gly Lys Val Arg Ala Val Val Lys Phe Asn Ala Ala Leu Ala 450 455 460 cac cac atc atg gcc ggc gcc gac gtg ctc gcc gtc acc agc cgc ttc 1440 His His Ile Met Ala Gly Ala Asp Val Leu Ala Val Thr Ser Arg Phe 465 470 475 480 gag ccc tgc ggc ctc atc cag ctg cag ggg atg cga tac gga acg ccc 1488 Glu Pro Cys Gly Leu Ile Gln Leu Gln Gly Met Arg Tyr Gly Thr Pro 485 490 495 tgc gcc tgc gcg tcc acc ggt gga ctc gtc gac acc atc atc gaa ggc 1536 Cys Ala Cys Ala Ser Thr Gly Gly Leu Val Asp Thr Ile Ile Glu Gly 500 505 510 aag acc ggg ttc cac atg ggc cgc ctc agc gtc gac tgt aac gtc gtg 1584 Lys Thr Gly Phe His Met Gly Arg Leu Ser Val Asp Cys Asn Val Val 515 520 525 gag ccg gcg gac gtc aag aag gtg gcc acc aca ttg cag cgc gcc atc 1632 Glu Pro Ala Asp Val Lys Lys Val Ala Thr Thr Leu Gln Arg Ala Ile 530 535 540 aag gtg gtc ggc acg ccg gcg tac gag gag atg gtg agg aac tgc atg 1680 Lys Val Val Gly Thr Pro Ala Tyr Glu Glu Met Val Arg Asn Cys Met 545 550 555 560 atc cag gat ctc tcc tgg aag ggc cct gcc aag aac tgg gag aac gtg 1728 Ile Gln Asp Leu Ser Trp Lys Gly Pro Ala Lys Asn Trp Glu Asn Val 565 570 575 ctg ctc agc ctc ggg gtc gcc ggc ggc gag cca ggg gtc gaa ggc gag 1776 Leu Leu Ser Leu Gly Val Ala Gly Gly Glu Pro Gly Val Glu Gly Glu 580 585 590 gag atc gcg ccg ctc gcc aag gag aac gtg gcc gcg ccc tga aga gtt 1824 Glu Ile Ala Pro Leu Ala Lys Glu Asn Val Ala Ala Pro Arg Val 595 600 605 cgg cct gca ggg ccc ctg atc tcg cgc gtg gtg caa aga tgt tgg gac 1872 Arg Pro Ala Gly Pro Leu Ile Ser Arg Val Val Gln Arg Cys Trp Asp 610 615 620 atc ttc tta tat atg ctg ttt cgt tta tgt gat atg gac aag t 1915 Ile Phe Leu Tyr Met Leu Phe Arg Leu Cys Asp Met Asp Lys 625 630 635 47 637 PRT Zea mays 47 Met Ala Ala Leu Ala Thr Ser Gln Leu Val Ala Thr Arg Ala Gly Leu 1 5 10 15 Gly Val Pro Asp Ala Ser Thr Phe Arg Arg Gly Ala Ala Gln Gly Leu 20 25 30 Arg Gly Ala Arg Ala Ser Ala Ala Ala Asp Thr Leu Ser Met Arg Thr 35 40 45 Ser Ala Arg Ala Ala Pro Arg His Gln Gln Gln Ala Ala Ala Pro Pro 50 55 60 Ser Lys Gly Ser Glu Gln His Thr Ala Ser Ala Gly Met Asn Val Val 65 70 75 80 Phe Val Gly Ala Glu Met Ala Pro Trp Ser Lys Thr Gly Gly Leu Gly 85 90 95 Asp Val Leu Gly Gly Leu Pro Pro Ala Met Ala Ala Asn Gly His Arg 100 105 110 Val Met Val Val Ser Pro Arg Tyr Asp Gln Tyr Lys Asp Ala Trp Asp 115 120 125 Thr Ser Val Val Ser Glu Ile Lys Met Gly Asp Gly Tyr Glu Thr Val 130 135 140 Arg Phe Phe His Cys Tyr Lys Arg Gly Val Asp Arg Val Phe Val Asp 145 150 155 160 His Pro Leu Phe Leu Glu Arg Val Trp Gly Lys Thr Glu Glu Lys Ile 165 170 175 Tyr Gly Pro Val Ala Gly Thr Asp Tyr Arg Asp Asn Gln Leu Arg Phe 180 185 190 Ser Leu Leu Cys Gln Gly Ala Leu Glu Ala Pro Arg Ile Leu Ser Leu 195 200 205 Asn Asn Asn Pro Tyr Phe Ser Gly Pro Tyr Gly Arg Asp Val Val Phe 210 215 220 Val Cys Asn Asp Trp His Thr Gly Pro Leu Ser Cys Tyr Leu Lys Ser 225 230 235 240 Asn Tyr Gln Ser His Gly Ile Tyr Arg Asp Ala Lys Thr Ala Phe Cys 245 250 255 Ile His Asn Ile Ser Tyr Gln Gly Arg Phe Ala Phe Ser Asp Tyr Pro 260 265 270 Glu Leu Asn Leu Pro Glu Arg Phe Lys Ser Ser Phe Asp Phe Ile Asp 275 280 285 Gly Tyr Glu Lys Pro Val Glu Gly Arg Lys Ile Asn Trp Met Lys Ala 290 295 300 Gly Ile Leu Glu Ala Asp Arg Val Leu Thr Val Ser Pro Tyr Tyr Ala 305 310 315 320 Glu Glu Leu Ile Ser Gly Ile Ala Arg Gly Cys Glu Leu Asp Asn Ile 325 330 335 Met Arg Leu Thr Gly Ile Thr Gly Ile Val Asn Gly Met Asp Val Ser 340 345 350 Glu Trp Asp Pro Ser Arg Asp Lys Tyr Ile Ala Val Lys Tyr Asp Val 355 360 365 Ser Thr Ala Val Glu Ala Lys Ala Leu Asn Lys Glu Ala Leu Gln Ala 370 375 380 Glu Val Gly Leu Pro Val Asp Arg Asn Ile Pro Leu Val Ala Phe Ile 385 390 395 400 Gly Arg Leu Glu Glu Gln Lys Gly Pro Asp Val Met Ala Ala Ala Ile 405 410 415 Pro Gln Leu Met Glu Met Val Glu Asp Val Gln Ile Val Leu Leu Gly 420 425 430 Thr Gly Lys Lys Lys Phe Glu Arg Met Leu Met Ser Ala Glu Glu Lys 435 440 445 Phe Pro Gly Lys Val Arg Ala Val Val Lys Phe Asn Ala Ala Leu Ala 450 455 460 His His Ile Met Ala Gly Ala Asp Val Leu Ala Val Thr Ser Arg Phe 465 470 475 480 Glu Pro Cys Gly Leu Ile Gln Leu Gln Gly Met Arg Tyr Gly Thr Pro 485 490 495 Cys Ala Cys Ala Ser Thr Gly Gly Leu Val Asp Thr Ile Ile Glu Gly 500 505 510 Lys Thr Gly Phe His Met Gly Arg Leu Ser Val Asp Cys Asn Val Val 515 520 525 Glu Pro Ala Asp Val Lys Lys Val Ala Thr Thr Leu Gln Arg Ala Ile 530 535 540 Lys Val Val Gly Thr Pro Ala Tyr Glu Glu Met Val Arg Asn Cys Met 545 550 555 560 Ile Gln Asp Leu Ser Trp Lys Gly Pro Ala Lys Asn Trp Glu Asn Val 565 570 575 Leu Leu Ser Leu Gly Val Ala Gly Gly Glu Pro Gly Val Glu Gly Glu 580 585 590 Glu Ile Ala Pro Leu Ala Lys Glu Asn Val Ala Ala Pro Arg Val Arg 595 600 605 Pro Ala Gly Pro Leu Ile Ser Arg Val Val Gln Arg Cys Trp Asp Ile 610 615 620 Phe Leu Tyr Met Leu Phe Arg Leu Cys Asp Met Asp Lys 625 630 635 48 2423 DNA Zea mays CDS (join(1..2094, 2098..2103, 2107..2304, 2308..2421)) 48 atg ccg ggg gca atc tct tcc tcg tcg tcg gct ttt ctc ctc ccc gtc 48 Met Pro Gly Ala Ile Ser Ser Ser Ser Ser Ala Phe Leu Leu Pro Val 1 5 10 15 gcg tcc tcc tcg ccg cgg cgc agg cgg ggc agt gtg ggt gct gct ctg 96 Ala Ser Ser Ser Pro Arg Arg Arg Arg Gly Ser Val Gly Ala Ala Leu 20 25 30 cgc tcg tac ggc tac agc ggc gcg gag ctg cgg ttg cat tgg gcg cgg 144 Arg Ser Tyr Gly Tyr Ser Gly Ala Glu Leu Arg Leu His Trp Ala Arg 35 40 45 cgg ggc ccg cct cag gat gga gcg gcg tcg gta cgc gcc gca gcg gca 192 Arg Gly Pro Pro Gln Asp Gly Ala Ala Ser Val Arg Ala Ala Ala Ala 50 55 60 ccg gcc ggg ggc gaa agc gag gag gca gcg aag agc tcc tcc tcg tcc 240 Pro Ala Gly Gly Glu Ser Glu Glu Ala Ala Lys Ser Ser Ser Ser Ser 65 70 75 80 cag gcg ggc gct gtt cag ggc agc acg gcc aag gct gtg gat tct gct 288 Gln Ala Gly Ala Val Gln Gly Ser Thr Ala Lys Ala Val Asp Ser Ala 85 90 95 tca cct ccc aat cct ttg aca tct gct ccg aag caa agt cag agc gct 336 Ser Pro Pro Asn Pro Leu Thr Ser Ala Pro Lys Gln Ser Gln Ser Ala 100 105 110 gca atg caa aac gga acg agt ggg ggc agc agc gcg agc acc gcc gcg 384 Ala Met Gln Asn Gly Thr Ser Gly Gly Ser Ser Ala Ser Thr Ala Ala 115 120 125 ccg gtg tcc gga ccc aaa gct gat cat cca tca gct cct gtc acc aag 432 Pro Val Ser Gly Pro Lys Ala Asp His Pro Ser Ala Pro Val Thr Lys 130 135 140 aga gaa atc gat gcc agt gcg gtg aag cca gag ccc gca ggt gat gat 480 Arg Glu Ile Asp Ala Ser Ala Val Lys Pro Glu Pro Ala Gly Asp Asp 145 150 155 160 gct aga ccg gtg gaa agc ata ggc atc gct gaa ccg gtg gat gct aag 528 Ala Arg Pro Val Glu Ser Ile Gly Ile Ala Glu Pro Val Asp Ala Lys 165 170 175 gct gat gca gct ccg gct aca gat gcg gcg gcg agt gct cct tat gac 576 Ala Asp Ala Ala Pro Ala Thr Asp Ala Ala Ala Ser Ala Pro Tyr Asp 180 185 190 agg gag gat aat gaa cct ggc cct ttg gct ggg cct aat gtg atg aac 624 Arg Glu Asp Asn Glu Pro Gly Pro Leu Ala Gly Pro Asn Val Met Asn 195 200 205 gtc gtc gtg gtg gct tct gaa tgt gct cct ttc tgc aag aca ggt ggc 672 Val Val Val Val Ala Ser Glu Cys Ala Pro Phe Cys Lys Thr Gly Gly 210 215 220 ctt gga gat gtc gtg ggt gct ttg cct aag gct ctg gcg agg aga gga 720 Leu Gly Asp Val Val Gly Ala Leu Pro Lys Ala Leu Ala Arg Arg Gly 225 230 235 240 cac cgt gtt atg gtc gtg ata cca aga tat gga gag tat gcc gaa gcc 768 His Arg Val Met Val Val Ile Pro Arg Tyr Gly Glu Tyr Ala Glu Ala 245 250 255 cgg gat tta ggt gta agg aga cgt tac aag gta gct gga cag gat tca 816 Arg Asp Leu Gly Val Arg Arg Arg Tyr Lys Val Ala Gly Gln Asp Ser 260 265 270 gaa gtt act tat ttt cac tct tac att gat gga gtt gat ttt gta ttc 864 Glu Val Thr Tyr Phe His Ser Tyr Ile Asp Gly Val Asp Phe Val Phe 275 280 285 gta gaa gcc cct ccc ttc cgg cac cgg cac aat aat att tat ggg gga 912 Val Glu Ala Pro Pro Phe Arg His Arg His Asn Asn Ile Tyr Gly Gly 290 295 300 gaa aga ttg gat att ttg aag cgc atg att ttg ttc tgc aag gcc gct 960 Glu Arg Leu Asp Ile Leu Lys Arg Met Ile Leu Phe Cys Lys Ala Ala 305 310 315 320 gtt gag gtt cca tgg tat gct cca tgt ggc ggt act gtc tat ggt gat 1008 Val Glu Val Pro Trp Tyr Ala Pro Cys Gly Gly Thr Val Tyr Gly Asp 325 330 335 ggc aac tta gtt ttc att gct aat gat tgg cat acc gca ctt ctg cct 1056 Gly Asn Leu Val Phe Ile Ala Asn Asp Trp His Thr Ala Leu Leu Pro 340 345 350 gtc tat cta aag gcc tat tac cgg gac aat ggt ttg atg cag tat gct 1104 Val Tyr Leu Lys Ala Tyr Tyr Arg Asp Asn Gly Leu Met Gln Tyr Ala 355 360 365 cgc tct gtg ctt gtg ata cac aac att gct cat cag ggt cgt ggc cct 1152 Arg Ser Val Leu Val Ile His Asn Ile Ala His Gln Gly Arg Gly Pro 370 375 380 gta gac gac ttc gtc aat ttt gac ttg cct gaa cac tac atc gac cac 1200 Val Asp Asp Phe Val Asn Phe Asp Leu Pro Glu His Tyr Ile Asp His 385 390 395 400 ttc aaa ctg tat gac aac att ggt ggg gat cac agc aac gtt ttt gct 1248 Phe Lys Leu Tyr Asp Asn Ile Gly Gly Asp His Ser Asn Val Phe Ala 405 410 415 gcg ggg ctg aag acg gca gac cgg gtg gtg acc gtt agc aat ggc tac 1296 Ala Gly Leu Lys Thr Ala Asp Arg Val Val Thr Val Ser Asn Gly Tyr 420 425 430 atg tgg gag ctg aag act tcg gaa ggc ggg tgg ggc ctc cac gac atc 1344 Met Trp Glu Leu Lys Thr Ser Glu Gly Gly Trp Gly Leu His Asp Ile 435 440 445 ata aac cag aac gac tgg aag ctg cag ggc atc gtg aac ggc atc gac 1392 Ile Asn Gln Asn Asp Trp Lys Leu Gln Gly Ile Val Asn Gly Ile Asp 450 455 460 atg agc gag tgg aac ccc gct gtg gac gtg cac ctc cac tcc gac gac 1440 Met Ser Glu Trp Asn Pro Ala Val Asp Val His Leu His Ser Asp Asp 465 470 475 480 tac acc aac tac acg ttc gag acg ctg gac acc ggc aag cgg cag tgc 1488 Tyr Thr Asn Tyr Thr Phe Glu Thr Leu Asp Thr Gly Lys Arg Gln Cys 485 490 495 aag gcc gcc ctg cag cgg cag ctg ggc ctg cag gtc cgc gac gac gtg 1536 Lys Ala Ala Leu Gln Arg Gln Leu Gly Leu Gln Val Arg Asp Asp Val 500 505 510 cca ctg atc ggg ttc atc ggg cgg ctg gac cac cag aag ggc gtg gac 1584 Pro Leu Ile Gly Phe Ile Gly Arg Leu Asp His Gln Lys Gly Val Asp 515 520 525 atc atc gcc gac gcg atc cac tgg atc gcg ggg cag gac gtg cag ctc 1632 Ile Ile Ala Asp Ala Ile His Trp Ile Ala Gly Gln Asp Val Gln Leu 530 535 540 gtg atg ctg ggc acc ggg cgg gcc gac ctg gag gac atg ctg cgg cgg 1680 Val Met Leu Gly Thr Gly Arg Ala Asp Leu Glu Asp Met Leu Arg Arg 545 550 555 560 ttc gag tcg gag cac agc gac aag gtg cgc gcg tgg gtg ggg ttc tcg 1728 Phe Glu Ser Glu His Ser Asp Lys Val Arg Ala Trp Val Gly Phe Ser 565 570 575 gtg ccc ctg gcg cac cgc atc acg gcg ggc gcg gac atc ctg ctg atg 1776 Val Pro Leu Ala His Arg Ile Thr Ala Gly Ala Asp Ile Leu Leu Met 580 585 590 ccg tcg cgg ttc gag ccg tgc ggg ctg aac cag ctc tac gcc atg gcg 1824 Pro Ser Arg Phe Glu Pro Cys Gly Leu Asn Gln Leu Tyr Ala Met Ala 595 600 605 tac ggg acc gtg ccc gtg gtg cac gcc gtg ggg ggg ctc cgg gac acg 1872 Tyr Gly Thr Val Pro Val Val His Ala Val Gly Gly Leu Arg Asp Thr 610 615 620 gtg gcg ccg ttc gac ccg ttc aac gac acc ggg ctc ggg tgg acg ttc 1920 Val Ala Pro Phe Asp Pro Phe Asn Asp Thr Gly Leu Gly Trp Thr Phe 625 630 635 640 gac cgc gcg gag gcg aac cgg atg atc gac gcg ctc tcg cac tgc ctc 1968 Asp Arg Ala Glu Ala Asn Arg Met Ile Asp Ala Leu Ser His Cys Leu 645 650 655 acc acg tac cgg aac tac aag gag agc tgg cgc gcc tgc agg gcg cgc 2016 Thr Thr Tyr Arg Asn Tyr Lys Glu Ser Trp Arg Ala Cys Arg Ala Arg 660 665 670 ggc atg gcc gag gac ctc agc tgg gac cac gcc gcc gtg ctg tat gag 2064 Gly Met Ala Glu Asp Leu Ser Trp Asp His Ala Ala Val Leu Tyr Glu 675 680 685 gac gtg ctc gtc aag gcg aag tac cag tgg tga gcg aat taa ttg gcg 2112 Asp Val Leu Val Lys Ala Lys Tyr Gln Trp Ala Asn Leu Ala 690 695 700 acg cga cgc cgc tcc tgt cgc agg acc tgg acg tta ttt aga agg ctc 2160 Thr Arg Arg Arg Ser Cys Arg Arg Thr Trp Thr Leu Phe Arg Arg Leu 705 710 715 ttc tcc ctg gcg gct ttg atg cgt gcg tcg cat ttg cgc cgg gcg gac 2208 Phe Ser Leu Ala Ala Leu Met Arg Ala Ser His Leu Arg Arg Ala Asp 720 725 730 ggg cga cgg tgg ttg gcc tac cgc cta cgt cgg ctg cgt gcc ctg gga 2256 Gly Arg Arg Trp Leu Ala Tyr Arg Leu Arg Arg Leu Arg Ala Leu Gly 735 740 745 750 att tgg gcg ggc acg atg atg cca ctg ggc acc ggg cgc ggg gta gta 2304 Ile Trp Ala Gly Thr Met Met Pro Leu Gly Thr Gly Arg Gly Val Val 755 760 765 tga tat gaa acc gac ggc gat gga gat gag gcg cat ggc att ttc cca 2352 Tyr Glu Thr Asp Gly Asp Gly Asp Glu Ala His Gly Ile Phe Pro 770 775 780 ctg ata aat ggg gag ttg tat gct act tta ata tcg cca ctc ctg tta 2400 Leu Ile Asn Gly Glu Leu Tyr Ala Thr Leu Ile Ser Pro Leu Leu Leu 785 790 795 gta ttt ata ttg atg gcg gcc gc 2423 Val Phe Ile Leu Met Ala Ala 800 49 804 PRT Zea mays 49 Met Pro Gly Ala Ile Ser Ser Ser Ser Ser Ala Phe Leu Leu Pro Val 1 5 10 15 Ala Ser Ser Ser Pro Arg Arg Arg Arg Gly Ser Val Gly Ala Ala Leu 20 25 30 Arg Ser Tyr Gly Tyr Ser Gly Ala Glu Leu Arg Leu His Trp Ala Arg 35 40 45 Arg Gly Pro Pro Gln Asp Gly Ala Ala Ser Val Arg Ala Ala Ala Ala 50 55 60 Pro Ala Gly Gly Glu Ser Glu Glu Ala Ala Lys Ser Ser Ser Ser Ser 65 70 75 80 Gln Ala Gly Ala Val Gln Gly Ser Thr Ala Lys Ala Val Asp Ser Ala 85 90 95 Ser Pro Pro Asn Pro Leu Thr Ser Ala Pro Lys Gln Ser Gln Ser Ala 100 105 110 Ala Met Gln Asn Gly Thr Ser Gly Gly Ser Ser Ala Ser Thr Ala Ala 115 120 125 Pro Val Ser Gly Pro Lys Ala Asp His Pro Ser Ala Pro Val Thr Lys 130 135 140 Arg Glu Ile Asp Ala Ser Ala Val Lys Pro Glu Pro Ala Gly Asp Asp 145 150 155 160 Ala Arg Pro Val Glu Ser Ile Gly Ile Ala Glu Pro Val Asp Ala Lys 165 170 175 Ala Asp Ala Ala Pro Ala Thr Asp Ala Ala Ala Ser Ala Pro Tyr Asp 180 185 190 Arg Glu Asp Asn Glu Pro Gly Pro Leu Ala Gly Pro Asn Val Met Asn 195 200 205 Val Val Val Val Ala Ser Glu Cys Ala Pro Phe Cys Lys Thr Gly Gly 210 215 220 Leu Gly Asp Val Val Gly Ala Leu Pro Lys Ala Leu Ala Arg Arg Gly 225 230 235 240 His Arg Val Met Val Val Ile Pro Arg Tyr Gly Glu Tyr Ala Glu Ala 245 250 255 Arg Asp Leu Gly Val Arg Arg Arg Tyr Lys Val Ala Gly Gln Asp Ser 260 265 270 Glu Val Thr Tyr Phe His Ser Tyr Ile Asp Gly Val Asp Phe Val Phe 275 280 285 Val Glu Ala Pro Pro Phe Arg His Arg His Asn Asn Ile Tyr Gly Gly 290 295 300 Glu Arg Leu Asp Ile Leu Lys Arg Met Ile Leu Phe Cys Lys Ala Ala 305 310 315 320 Val Glu Val Pro Trp Tyr Ala Pro Cys Gly Gly Thr Val Tyr Gly Asp 325 330 335 Gly Asn Leu Val Phe Ile Ala Asn Asp Trp His Thr Ala Leu Leu Pro 340 345 350 Val Tyr Leu Lys Ala Tyr Tyr Arg Asp Asn Gly Leu Met Gln Tyr Ala 355 360 365 Arg Ser Val Leu Val Ile His Asn Ile Ala His Gln Gly Arg Gly Pro 370 375 380 Val Asp Asp Phe Val Asn Phe Asp Leu Pro Glu His Tyr Ile Asp His 385 390 395 400 Phe Lys Leu Tyr Asp Asn Ile Gly Gly Asp His Ser Asn Val Phe Ala 405 410 415 Ala Gly Leu Lys Thr Ala Asp Arg Val Val Thr Val Ser Asn Gly Tyr 420 425 430 Met Trp Glu Leu Lys Thr Ser Glu Gly Gly Trp Gly Leu His Asp Ile 435 440 445 Ile Asn Gln Asn Asp Trp Lys Leu Gln Gly Ile Val Asn Gly Ile Asp 450 455 460 Met Ser Glu Trp Asn Pro Ala Val Asp Val His Leu His Ser Asp Asp 465 470 475 480 Tyr Thr Asn Tyr Thr Phe Glu Thr Leu Asp Thr Gly Lys Arg Gln Cys 485 490 495 Lys Ala Ala Leu Gln Arg Gln Leu Gly Leu Gln Val Arg Asp Asp Val 500 505 510 Pro Leu Ile Gly Phe Ile Gly Arg Leu Asp His Gln Lys Gly Val Asp 515 520 525 Ile Ile Ala Asp Ala Ile His Trp Ile Ala Gly Gln Asp Val Gln Leu 530 535 540 Val Met Leu Gly Thr Gly Arg Ala Asp Leu Glu Asp Met Leu Arg Arg 545 550 555 560 Phe Glu Ser Glu His Ser Asp Lys Val Arg Ala Trp Val Gly Phe Ser 565 570 575 Val Pro Leu Ala His Arg Ile Thr Ala Gly Ala Asp Ile Leu Leu Met 580 585 590 Pro Ser Arg Phe Glu Pro Cys Gly Leu Asn Gln Leu Tyr Ala Met Ala 595 600 605 Tyr Gly Thr Val Pro Val Val His Ala Val Gly Gly Leu Arg Asp Thr 610 615 620 Val Ala Pro Phe Asp Pro Phe Asn Asp Thr Gly Leu Gly Trp Thr Phe 625 630 635 640 Asp Arg Ala Glu Ala Asn Arg Met Ile Asp Ala Leu Ser His Cys Leu 645 650 655 Thr Thr Tyr Arg Asn Tyr Lys Glu Ser Trp Arg Ala Cys Arg Ala Arg 660 665 670 Gly Met Ala Glu Asp Leu Ser Trp Asp His Ala Ala Val Leu Tyr Glu 675 680 685 Asp Val Leu Val Lys Ala Lys Tyr Gln Trp Ala Asn Leu Ala Thr Arg 690 695 700 Arg Arg Ser Cys Arg Arg Thr Trp Thr Leu Phe Arg Arg Leu Phe Ser 705 710 715 720 Leu Ala Ala Leu Met Arg Ala Ser His Leu Arg Arg Ala Asp Gly Arg 725 730 735 Arg Trp Leu Ala Tyr Arg Leu Arg Arg Leu Arg Ala Leu Gly Ile Trp 740 745 750 Ala Gly Thr Met Met Pro Leu Gly Thr Gly Arg Gly Val Val Tyr Glu 755 760 765 Thr Asp Gly Asp Gly Asp Glu Ala His Gly Ile Phe Pro Leu Ile Asn 770 775 780 Gly Glu Leu Tyr Ala Thr Leu Ile Ser Pro Leu Leu Leu Val Phe Ile 785 790 795 800 Leu Met Ala Ala 50 2010 DNA Zea mays CDS (1)..(2010) 50 gct gag gct gag gcc ggg ggc aag gac gcg ccg ccg gag agg agc ggc 48 Ala Glu Ala Glu Ala Gly Gly Lys Asp Ala Pro Pro Glu Arg Ser Gly 1 5 10 15 gac gcc gcc agg ttg ccc cgc gct cgg cgc aat gcg gtc tcc aaa cgg 96 Asp Ala Ala Arg Leu Pro Arg Ala Arg Arg Asn Ala Val Ser Lys Arg 20 25 30 agg gat cct ctt cag ccg gtc ggc cgg tac ggc tcc gcg acg gga aac 144 Arg Asp Pro Leu Gln Pro Val Gly Arg Tyr Gly Ser Ala Thr Gly Asn 35 40 45 acg gcc agg acc ggc gcc gcg tcc tgc cag aac gcc gca ttg gcg gac 192 Thr Ala Arg Thr Gly Ala Ala Ser Cys Gln Asn Ala Ala Leu Ala Asp 50 55 60 gtt gag atc aag tcc atc gtc gcc gcg ccg ccg acg agc ata gtg aag 240 Val Glu Ile Lys Ser Ile Val Ala Ala Pro Pro Thr Ser Ile Val Lys 65 70 75 80 ttc cca gcg ccg ggc tac agg atg atc ctt ccc tct ggg gac ata gcg 288 Phe Pro Ala Pro Gly Tyr Arg Met Ile Leu Pro Ser Gly Asp Ile Ala 85 90 95 ccg gag act gtc ctc cca gcc ccg aag cca ctg cat gaa tcg cct gcg 336 Pro Glu Thr Val Leu Pro Ala Pro Lys Pro Leu His Glu Ser Pro Ala 100 105 110 gtt gac gga gat tca aat gga att gca cct cct aca gtt gag cca tta 384 Val Asp Gly Asp Ser Asn Gly Ile Ala Pro Pro Thr Val Glu Pro Leu 115 120 125 gta cag gag gcc act tgg gat ttc aag aaa tac atc ggt ttt gac gag 432 Val Gln Glu Ala Thr Trp Asp Phe Lys Lys Tyr Ile Gly Phe Asp Glu 130 135 140 cct gac gaa gcg aag gat gat tcc agg gtt ggt gca gat gat gct ggt 480 Pro Asp Glu Ala Lys Asp Asp Ser Arg Val Gly Ala Asp Asp Ala Gly 145 150 155 160 tct ttt gaa cat tat ggg gac aat gat tct ggg cct ttg gcc ggg gag 528 Ser Phe Glu His Tyr Gly Asp Asn Asp Ser Gly Pro Leu Ala Gly Glu 165 170 175 aat gtt atg aac gtg atc gtg gtg gct gct gaa tgt tct cca tgg tgc 576 Asn Val Met Asn Val Ile Val Val Ala Ala Glu Cys Ser Pro Trp Cys 180 185 190 aaa aca ggt ggt ctt gga gat gtt gtg gga gct tta ccc aag gct tta 624 Lys Thr Gly Gly Leu Gly Asp Val Val Gly Ala Leu Pro Lys Ala Leu 195 200 205 gcg aga aga gga cat cgt gtt atg gtt gtg gta cca agg tat ggg gac 672 Ala Arg Arg Gly His Arg Val Met Val Val Val Pro Arg Tyr Gly Asp 210 215 220 tat gtg gaa gcc ttt gat atg gga atc cgg aaa tac tac aaa gct gca 720 Tyr Val Glu Ala Phe Asp Met Gly Ile Arg Lys Tyr Tyr Lys Ala Ala 225 230 235 240 gga cag gac cta gaa gtg aac tat ttc cat gca ttt att gat gga gtc 768 Gly Gln Asp Leu Glu Val Asn Tyr Phe His Ala Phe Ile Asp Gly Val 245 250 255 gac ttt gtg ttc att gat gcc cct ctt ttc cgg cac cgt caa gat gac 816 Asp Phe Val Phe Ile Asp Ala Pro Leu Phe Arg His Arg Gln Asp Asp 260 265 270 ata tat ggg gga agt agg cag gaa atc atg aag cgc atg att ttg ttt 864 Ile Tyr Gly Gly Ser Arg Gln Glu Ile Met Lys Arg Met Ile Leu Phe 275 280 285 tgc aag gtt gct gtt gag gtt cct tgg cac gtt cca tgc ggt ggt gtg 912 Cys Lys Val Ala Val Glu Val Pro Trp His Val Pro Cys Gly Gly Val 290 295 300 tgc tac gga gat gga aat ttg gtg ttc att gcc aat gat tgg cac act 960 Cys Tyr Gly Asp Gly Asn Leu Val Phe Ile Ala Asn Asp Trp His Thr 305 310 315 320 gca ctc ctg cct gtt tat ctg aag gca tat tac aga gac cat ggg tta 1008 Ala Leu Leu Pro Val Tyr Leu Lys Ala Tyr Tyr Arg Asp His Gly Leu 325 330 335 atg cag tac act cgc tcc gtc ctc gtc ata cat aac atc gcc cac cag 1056 Met Gln Tyr Thr Arg Ser Val Leu Val Ile His Asn Ile Ala His Gln 340 345 350 ggc cgt ggt cct gta gat gaa ttc ccg tac atg gac ttg cct gaa cac 1104 Gly Arg Gly Pro Val Asp Glu Phe Pro Tyr Met Asp Leu Pro Glu His 355 360 365 tac ctt caa cat ttc gag ctg tac gat ccc gtc ggt ggc gag cac gcc 1152 Tyr Leu Gln His Phe Glu Leu Tyr Asp Pro Val Gly Gly Glu His Ala 370 375 380 aac atc ttt gcc gcg ggt ctg aag atg gca gac cgg gtg gtg act gtc 1200 Asn Ile Phe Ala Ala Gly Leu Lys Met Ala Asp Arg Val Val Thr Val 385 390 395 400 agc cgc ggc tac ctg tgg gag ctg aag aca gtg gaa ggc ggc tgg ggc 1248 Ser Arg Gly Tyr Leu Trp Glu Leu Lys Thr Val Glu Gly Gly Trp Gly 405 410 415 ctc cac gac atc atc cgt tct aac gac tgg aag atc aat ggc atc gtg 1296 Leu His Asp Ile Ile Arg Ser Asn Asp Trp Lys Ile Asn Gly Ile Val 420 425 430 aac ggc atc gac cac cag gag tgg aac ccc aag gtg gac gtg cac ctg 1344 Asn Gly Ile Asp His Gln Glu Trp Asn Pro Lys Val Asp Val His Leu 435 440 445 cgg tcg gac ggc tac acc aac tac tcc ctc gag aca ctc gac gct gga 1392 Arg Ser Asp Gly Tyr Thr Asn Tyr Ser Leu Glu Thr Leu Asp Ala Gly 450 455 460 aag cgg cag tgc aag gcg gcc ctg cag cgg gag ctg ggc ctg gaa gtg 1440 Lys Arg Gln Cys Lys Ala Ala Leu Gln Arg Glu Leu Gly Leu Glu Val 465 470 475 480 cgc gac gac gtg ccg ctg ctc ggc ttc atc ggg cgt ctg gat gga cag 1488 Arg Asp Asp Val Pro Leu Leu Gly Phe Ile Gly Arg Leu Asp Gly Gln 485 490 495 aag ggc gtg gac atc atc ggg gac gcg atg ccg tgg atc gcg ggg cag 1536 Lys Gly Val Asp Ile Ile Gly Asp Ala Met Pro Trp Ile Ala Gly Gln 500 505 510 gac gtg cag ctg gtg atg ctg ggc acc ggg cgc gcc gac ctg gaa cga 1584 Asp Val Gln Leu Val Met Leu Gly Thr Gly Arg Ala Asp Leu Glu Arg 515 520 525 atg ctg cag cac ttg gag cgg gag cat ccc aac aag gtg cgc ggg tgg 1632 Met Leu Gln His Leu Glu Arg Glu His Pro Asn Lys Val Arg Gly Trp 530 535 540 gtc ggg ttc tcg gtg cct atg gcg cat cgc atc acg gcg ggc gcc gac 1680 Val Gly Phe Ser Val Pro Met Ala His Arg Ile Thr Ala Gly Ala Asp 545 550 555 560 gtg ctg gtg atg cct tcc cgc ttc gag ccc tgc ggg ctg aac cag ctc 1728 Val Leu Val Met Pro Ser Arg Phe Glu Pro Cys Gly Leu Asn Gln Leu 565 570 575 tac gcg atg gcg tac ggc acc gtc cct gtg gtg cac gcc gtg ggc ggg 1776 Tyr Ala Met Ala Tyr Gly Thr Val Pro Val Val His Ala Val Gly Gly 580 585 590 ctc agg gac acc gtg gcg ccg ttc gac ccg ttc ggc gac gcc ggg ctc 1824 Leu Arg Asp Thr Val Ala Pro Phe Asp Pro Phe Gly Asp Ala Gly Leu 595 600 605 ggg tgg act ttt gac cgc gcc gag gcc aac aag ctg atc gag gcg ctc 1872 Gly Trp Thr Phe Asp Arg Ala Glu Ala Asn Lys Leu Ile Glu Ala Leu 610 615 620 agg cac tgc ctc gac acg tac cgg aag tac ggg gag agc tgg aag agt 1920 Arg His Cys Leu Asp Thr Tyr Arg Lys Tyr Gly Glu Ser Trp Lys Ser 625 630 635 640 ctc cag gcg cgc ggc atg tcg cag gac ctc agc tgg gac cac gcg gct 1968 Leu Gln Ala Arg Gly Met Ser Gln Asp Leu Ser Trp Asp His Ala Ala 645 650 655 gag ctc tac gag gac gtc ctt gtc aag gcc aag tac cag tgg 2010 Glu Leu Tyr Glu Asp Val Leu Val Lys Ala Lys Tyr Gln Trp 660 665 670 51 670 PRT Zea mays 51 Ala Glu Ala Glu Ala Gly Gly Lys Asp Ala Pro Pro Glu Arg Ser Gly 1 5 10 15 Asp Ala Ala Arg Leu Pro Arg Ala Arg Arg Asn Ala Val Ser Lys Arg 20 25 30 Arg Asp Pro Leu Gln Pro Val Gly Arg Tyr Gly Ser Ala Thr Gly Asn 35 40 45 Thr Ala Arg Thr Gly Ala Ala Ser Cys Gln Asn Ala Ala Leu Ala Asp 50 55 60 Val Glu Ile Lys Ser Ile Val Ala Ala Pro Pro Thr Ser Ile Val Lys 65 70 75 80 Phe Pro Ala Pro Gly Tyr Arg Met Ile Leu Pro Ser Gly Asp Ile Ala 85 90 95 Pro Glu Thr Val Leu Pro Ala Pro Lys Pro Leu His Glu Ser Pro Ala 100 105 110 Val Asp Gly Asp Ser Asn Gly Ile Ala Pro Pro Thr Val Glu Pro Leu 115 120 125 Val Gln Glu Ala Thr Trp Asp Phe Lys Lys Tyr Ile Gly Phe Asp Glu 130 135 140 Pro Asp Glu Ala Lys Asp Asp Ser Arg Val Gly Ala Asp Asp Ala Gly 145 150 155 160 Ser Phe Glu His Tyr Gly Asp Asn Asp Ser Gly Pro Leu Ala Gly Glu 165 170 175 Asn Val Met Asn Val Ile Val Val Ala Ala Glu Cys Ser Pro Trp Cys 180 185 190 Lys Thr Gly Gly Leu Gly Asp Val Val Gly Ala Leu Pro Lys Ala Leu 195 200 205 Ala Arg Arg Gly His Arg Val Met Val Val Val Pro Arg Tyr Gly Asp 210 215 220 Tyr Val Glu Ala Phe Asp Met Gly Ile Arg Lys Tyr Tyr Lys Ala Ala 225 230 235 240 Gly Gln Asp Leu Glu Val Asn Tyr Phe His Ala Phe Ile Asp Gly Val 245 250 255 Asp Phe Val Phe Ile Asp Ala Pro Leu Phe Arg His Arg Gln Asp Asp 260 265 270 Ile Tyr Gly Gly Ser Arg Gln Glu Ile Met Lys Arg Met Ile Leu Phe 275 280 285 Cys Lys Val Ala Val Glu Val Pro Trp His Val Pro Cys Gly Gly Val 290 295 300 Cys Tyr Gly Asp Gly Asn Leu Val Phe Ile Ala Asn Asp Trp His Thr 305 310 315 320 Ala Leu Leu Pro Val Tyr Leu Lys Ala Tyr Tyr Arg Asp His Gly Leu 325 330 335 Met Gln Tyr Thr Arg Ser Val Leu Val Ile His Asn Ile Ala His Gln 340 345 350 Gly Arg Gly Pro Val Asp Glu Phe Pro Tyr Met Asp Leu Pro Glu His 355 360 365 Tyr Leu Gln His Phe Glu Leu Tyr Asp Pro Val Gly Gly Glu His Ala 370 375 380 Asn Ile Phe Ala Ala Gly Leu Lys Met Ala Asp Arg Val Val Thr Val 385 390 395 400 Ser Arg Gly Tyr Leu Trp Glu Leu Lys Thr Val Glu Gly Gly Trp Gly 405 410 415 Leu His Asp Ile Ile Arg Ser Asn Asp Trp Lys Ile Asn Gly Ile Val 420 425 430 Asn Gly Ile Asp His Gln Glu Trp Asn Pro Lys Val Asp Val His Leu 435 440 445 Arg Ser Asp Gly Tyr Thr Asn Tyr Ser Leu Glu Thr Leu Asp Ala Gly 450 455 460 Lys Arg Gln Cys Lys Ala Ala Leu Gln Arg Glu Leu Gly Leu Glu Val 465 470 475 480 Arg Asp Asp Val Pro Leu Leu Gly Phe Ile Gly Arg Leu Asp Gly Gln 485 490 495 Lys Gly Val Asp Ile Ile Gly Asp Ala Met Pro Trp Ile Ala Gly Gln 500 505 510 Asp Val Gln Leu Val Met Leu Gly Thr Gly Arg Ala Asp Leu Glu Arg 515 520 525 Met Leu Gln His Leu Glu Arg Glu His Pro Asn Lys Val Arg Gly Trp 530 535 540 Val Gly Phe Ser Val Pro Met Ala His Arg Ile Thr Ala Gly Ala Asp 545 550 555 560 Val Leu Val Met Pro Ser Arg Phe Glu Pro Cys Gly Leu Asn Gln Leu 565 570 575 Tyr Ala Met Ala Tyr Gly Thr Val Pro Val Val His Ala Val Gly Gly 580 585 590 Leu Arg Asp Thr Val Ala Pro Phe Asp Pro Phe Gly Asp Ala Gly Leu 595 600 605 Gly Trp Thr Phe Asp Arg Ala Glu Ala Asn Lys Leu Ile Glu Ala Leu 610 615 620 Arg His Cys Leu Asp Thr Tyr Arg Lys Tyr Gly Glu Ser Trp Lys Ser 625 630 635 640 Leu Gln Ala Arg Gly Met Ser Gln Asp Leu Ser Trp Asp His Ala Ala 645 650 655 Glu Leu Tyr Glu Asp Val Leu Val Lys Ala Lys Tyr Gln Trp 660 665 670 52 1749 DNA Zea mays CDS (1)..(1749) 52 tgc gtc gcg gag ctg agc agg gag ggg ccc gcg ccg cgc ccg ctg cca 48 Cys Val Ala Glu Leu Ser Arg Glu Gly Pro Ala Pro Arg Pro Leu Pro 1 5 10 15 ccc gcg ctg ctg gcg ccc ccg ctc gtg ccc ggc ttc ctc gcg ccg ccg 96 Pro Ala Leu Leu Ala Pro Pro Leu Val Pro Gly Phe Leu Ala Pro Pro 20 25 30 gcc gag ccc acg ggt gag ccg gca tcg acg ccg ccg ccc gtg ccc gac 144 Ala Glu Pro Thr Gly Glu Pro Ala Ser Thr Pro Pro Pro Val Pro Asp 35 40 45 gcc ggc ctg ggg gac ctc ggt ctc gaa cct gaa ggg att gct gaa ggt 192 Ala Gly Leu Gly Asp Leu Gly Leu Glu Pro Glu Gly Ile Ala Glu Gly 50 55 60 tcc atc gat aac aca gta gtt gtg gca agt gag caa gat tct gag att 240 Ser Ile Asp Asn Thr Val Val Val Ala Ser Glu Gln Asp Ser Glu Ile 65 70 75 80 gtg gtt gga aag gag caa gct cga gct aaa gta aca caa agc att gtc 288 Val Val Gly Lys Glu Gln Ala Arg Ala Lys Val Thr Gln Ser Ile Val 85 90 95 ttt gta acc ggc gaa gct tct cct tat gca aag tct ggg ggt cta gga 336 Phe Val Thr Gly Glu Ala Ser Pro Tyr Ala Lys Ser Gly Gly Leu Gly 100 105 110 gat gtt tgt ggt tca ttg cca gtt gct ctt gct gct cgt ggt cac cgt 384 Asp Val Cys Gly Ser Leu Pro Val Ala Leu Ala Ala Arg Gly His Arg 115 120 125 gtg atg gtt gta atg ccc aga tat tta aat ggt acc tcc gat aag aat 432 Val Met Val Val Met Pro Arg Tyr Leu Asn Gly Thr Ser Asp Lys Asn 130 135 140 tat gca aat gca ttt tac aca gaa aaa cac att cgg att cca tgc ttt 480 Tyr Ala Asn Ala Phe Tyr Thr Glu Lys His Ile Arg Ile Pro Cys Phe 145 150 155 160 ggc ggt gaa cat gaa gtt acc ttc ttc cat gag tat aga gat tca gtt 528 Gly Gly Glu His Glu Val Thr Phe Phe His Glu Tyr Arg Asp Ser Val 165 170 175 gac tgg gtg ttt gtt gat cat ccc tca tat cac aga cct gga aat tta 576 Asp Trp Val Phe Val Asp His Pro Ser Tyr His Arg Pro Gly Asn Leu 180 185 190 tat gga gat aag ttt ggt gct ttt ggt gat aat cag ttc aga tac aca 624 Tyr Gly Asp Lys Phe Gly Ala Phe Gly Asp Asn Gln Phe Arg Tyr Thr 195 200 205 ctc ctt tgc tat gct gca tgt gag gct cct ttg atc ctt gaa ttg gga 672 Leu Leu Cys Tyr Ala Ala Cys Glu Ala Pro Leu Ile Leu Glu Leu Gly 210 215 220 gga tat att tat gga cag aat tgc atg ttt gtt gtc aat gat tgg cat 720 Gly Tyr Ile Tyr Gly Gln Asn Cys Met Phe Val Val Asn Asp Trp His 225 230 235 240 gcc agt cta gtg cca gtc ctt ctt gct gca aaa tat aga cca tat ggt 768 Ala Ser Leu Val Pro Val Leu Leu Ala Ala Lys Tyr Arg Pro Tyr Gly 245 250 255 gtt tat aaa gac tcc cgc agc att ctt gta ata cat aat tta gca cat 816 Val Tyr Lys Asp Ser Arg Ser Ile Leu Val Ile His Asn Leu Ala His 260 265 270 cag ggt gta gag cct gca agc aca tat cct gac ctt ggg ttg cca cct 864 Gln Gly Val Glu Pro Ala Ser Thr Tyr Pro Asp Leu Gly Leu Pro Pro 275 280 285 gaa tgg tat gga gct ctg gag tgg gta ttc cct gaa tgg gcg agg agg 912 Glu Trp Tyr Gly Ala Leu Glu Trp Val Phe Pro Glu Trp Ala Arg Arg 290 295 300 cat gcc ctt gac aag ggt gag gca gtt aat ttt ttg aaa ggt gca gtt 960 His Ala Leu Asp Lys Gly Glu Ala Val Asn Phe Leu Lys Gly Ala Val 305 310 315 320 gtg aca gca gat cga atc gtg act gtc agt aag ggt tat tcg tgg gag 1008 Val Thr Ala Asp Arg Ile Val Thr Val Ser Lys Gly Tyr Ser Trp Glu 325 330 335 gtc aca act gct gaa ggt gga cag ggc ctc aat gag ctc tta agc tcc 1056 Val Thr Thr Ala Glu Gly Gly Gln Gly Leu Asn Glu Leu Leu Ser Ser 340 345 350 aga aag agt gta tta aac gga att gta aat gga att gac att aat gat 1104 Arg Lys Ser Val Leu Asn Gly Ile Val Asn Gly Ile Asp Ile Asn Asp 355 360 365 tgg aac cct gcc aca gac aaa tgt atc ccc tgt cat tat tct gtt gat 1152 Trp Asn Pro Ala Thr Asp Lys Cys Ile Pro Cys His Tyr Ser Val Asp 370 375 380 gac ctc tct gga aag gcc aaa tgt aaa ggt gca ttg cag aag gag ctg 1200 Asp Leu Ser Gly Lys Ala Lys Cys Lys Gly Ala Leu Gln Lys Glu Leu 385 390 395 400 ggt tta cct ata agg cct gat gtt cct ctg att ggc ttt att gga agg 1248 Gly Leu Pro Ile Arg Pro Asp Val Pro Leu Ile Gly Phe Ile Gly Arg 405 410 415 ttg gat tat cag aaa ggc att gat ctc att caa ctt atc ata cca gat 1296 Leu Asp Tyr Gln Lys Gly Ile Asp Leu Ile Gln Leu Ile Ile Pro Asp 420 425 430 ctc atg cgg gaa gat gtt caa ttt gtc atg ctt gga tct ggt gac cca 1344 Leu Met Arg Glu Asp Val Gln Phe Val Met Leu Gly Ser Gly Asp Pro 435 440 445 gag ctt gaa gat tgg atg aga tct aca gag tcg atc ttc aag gat aaa 1392 Glu Leu Glu Asp Trp Met Arg Ser Thr Glu Ser Ile Phe Lys Asp Lys 450 455 460 ttt cgt gga tgg gtt gga ttt agt gtt cca gtt tcc cac cga ata act 1440 Phe Arg Gly Trp Val Gly Phe Ser Val Pro Val Ser His Arg Ile Thr 465 470 475 480 gcc ggc tgc gat ata ttg tta atg cca tcc aga ttc gaa cct tgt ggt 1488 Ala Gly Cys Asp Ile Leu Leu Met Pro Ser Arg Phe Glu Pro Cys Gly 485 490 495 ctc aat cag cta tat gct atg cag tat ggc aca gtt cct gtt gtc cat 1536 Leu Asn Gln Leu Tyr Ala Met Gln Tyr Gly Thr Val Pro Val Val His 500 505 510 gca act ggg ggc ctt aga gat acc gtg gag aac ttc aac cct ttc ggt 1584 Ala Thr Gly Gly Leu Arg Asp Thr Val Glu Asn Phe Asn Pro Phe Gly 515 520 525 gag aat gga gag cag ggt aca ggg tgg gca ttc gca ccc cta acc aca 1632 Glu Asn Gly Glu Gln Gly Thr Gly Trp Ala Phe Ala Pro Leu Thr Thr 530 535 540 gaa aac atg ttt gtg gac att gcg aac tgc aat atc tac ata cag gga 1680 Glu Asn Met Phe Val Asp Ile Ala Asn Cys Asn Ile Tyr Ile Gln Gly 545 550 555 560 aca caa gtc ctc ctg gga agg gct aat gaa gcg agg cat gtc aaa aga 1728 Thr Gln Val Leu Leu Gly Arg Ala Asn Glu Ala Arg His Val Lys Arg 565 570 575 ctt cac gtg gga cca tgc cgc 1749 Leu His Val Gly Pro Cys Arg 580 53 583 PRT Zea mays 53 Cys Val Ala Glu Leu Ser Arg Glu Gly Pro Ala Pro Arg Pro Leu Pro 1 5 10 15 Pro Ala Leu Leu Ala Pro Pro Leu Val Pro Gly Phe Leu Ala Pro Pro 20 25 30 Ala Glu Pro Thr Gly Glu Pro Ala Ser Thr Pro Pro Pro Val Pro Asp 35 40 45 Ala Gly Leu Gly Asp Leu Gly Leu Glu Pro Glu Gly Ile Ala Glu Gly 50 55 60 Ser Ile Asp Asn Thr Val Val Val Ala Ser Glu Gln Asp Ser Glu Ile 65 70 75 80 Val Val Gly Lys Glu Gln Ala Arg Ala Lys Val Thr Gln Ser Ile Val 85 90 95 Phe Val Thr Gly Glu Ala Ser Pro Tyr Ala Lys Ser Gly Gly Leu Gly 100 105 110 Asp Val Cys Gly Ser Leu Pro Val Ala Leu Ala Ala Arg Gly His Arg 115 120 125 Val Met Val Val Met Pro Arg Tyr Leu Asn Gly Thr Ser Asp Lys Asn 130 135 140 Tyr Ala Asn Ala Phe Tyr Thr Glu Lys His Ile Arg Ile Pro Cys Phe 145 150 155 160 Gly Gly Glu His Glu Val Thr Phe Phe His Glu Tyr Arg Asp Ser Val 165 170 175 Asp Trp Val Phe Val Asp His Pro Ser Tyr His Arg Pro Gly Asn Leu 180 185 190 Tyr Gly Asp Lys Phe Gly Ala Phe Gly Asp Asn Gln Phe Arg Tyr Thr 195 200 205 Leu Leu Cys Tyr Ala Ala Cys Glu Ala Pro Leu Ile Leu Glu Leu Gly 210 215 220 Gly Tyr Ile Tyr Gly Gln Asn Cys Met Phe Val Val Asn Asp Trp His 225 230 235 240 Ala Ser Leu Val Pro Val Leu Leu Ala Ala Lys Tyr Arg Pro Tyr Gly 245 250 255 Val Tyr Lys Asp Ser Arg Ser Ile Leu Val Ile His Asn Leu Ala His 260 265 270 Gln Gly Val Glu Pro Ala Ser Thr Tyr Pro Asp Leu Gly Leu Pro Pro 275 280 285 Glu Trp Tyr Gly Ala Leu Glu Trp Val Phe Pro Glu Trp Ala Arg Arg 290 295 300 His Ala Leu Asp Lys Gly Glu Ala Val Asn Phe Leu Lys Gly Ala Val 305 310 315 320 Val Thr Ala Asp Arg Ile Val Thr Val Ser Lys Gly Tyr Ser Trp Glu 325 330 335 Val Thr Thr Ala Glu Gly Gly Gln Gly Leu Asn Glu Leu Leu Ser Ser 340 345 350 Arg Lys Ser Val Leu Asn Gly Ile Val Asn Gly Ile Asp Ile Asn Asp 355 360 365 Trp Asn Pro Ala Thr Asp Lys Cys Ile Pro Cys His Tyr Ser Val Asp 370 375 380 Asp Leu Ser Gly Lys Ala Lys Cys Lys Gly Ala Leu Gln Lys Glu Leu 385 390 395 400 Gly Leu Pro Ile Arg Pro Asp Val Pro Leu Ile Gly Phe Ile Gly Arg 405 410 415 Leu Asp Tyr Gln Lys Gly Ile Asp Leu Ile Gln Leu Ile Ile Pro Asp 420 425 430 Leu Met Arg Glu Asp Val Gln Phe Val Met Leu Gly Ser Gly Asp Pro 435 440 445 Glu Leu Glu Asp Trp Met Arg Ser Thr Glu Ser Ile Phe Lys Asp Lys 450 455 460 Phe Arg Gly Trp Val Gly Phe Ser Val Pro Val Ser His Arg Ile Thr 465 470 475 480 Ala Gly Cys Asp Ile Leu Leu Met Pro Ser Arg Phe Glu Pro Cys Gly 485 490 495 Leu Asn Gln Leu Tyr Ala Met Gln Tyr Gly Thr Val Pro Val Val His 500 505 510 Ala Thr Gly Gly Leu Arg Asp Thr Val Glu Asn Phe Asn Pro Phe Gly 515 520 525 Glu Asn Gly Glu Gln Gly Thr Gly Trp Ala Phe Ala Pro Leu Thr Thr 530 535 540 Glu Asn Met Phe Val Asp Ile Ala Asn Cys Asn Ile Tyr Ile Gln Gly 545 550 555 560 Thr Gln Val Leu Leu Gly Arg Ala Asn Glu Ala Arg His Val Lys Arg 565 570 575 Leu His Val Gly Pro Cys Arg 580 54 870 PRT Zea mays 54 Met Ala Phe Arg Val Ser Gly Ala Val Leu Gly Gly Ala Val Arg Ala 1 5 10 15 Pro Arg Leu Thr Gly Gly Gly Glu Gly Ser Leu Val Phe Arg His Thr 20 25 30 Gly Leu Phe Leu Thr Arg Gly Ala Arg Val Gly Cys Ser Gly Thr His 35 40 45 Gly Ala Met Arg Ala Ala Ala Ala Ala Arg Lys Ala Met Val Pro Glu 50 55 60 Gly Glu Asn Asp Gly Leu Ala Ser Arg Ala Asp Ser Ala Gln Phe Gln 65 70 75 80 Ser Asp Glu Leu Glu Val Pro Asp Ile Ser Glu Glu Thr Thr Cys Gly 85 90 95 Ala Gly Val Ala Asp Ala Gln Ala Leu Asn Arg Val Arg Val Val Pro 100 105 110 Pro Pro Ser Asp Gly Gln Lys Ile Phe Gln Ile Asp Pro Met Leu Gln 115 120 125 Gly Tyr Lys Tyr His Leu Glu Tyr Arg Tyr Ser Leu Tyr Arg Arg Ile 130 135 140 Arg Ser Asp Ile Asp Glu His Glu Gly Gly Leu Glu Ala Phe Ser Arg 145 150 155 160 Ser Tyr Glu Lys Phe Gly Phe Asn Ala Ser Ala Glu Gly Ile Thr Tyr 165 170 175 Arg Glu Trp Ala Pro Gly Ala Phe Ser Ala Ala Leu Val Gly Asp Val 180 185 190 Asn Asn Trp Asp Pro Asn Ala Asp Arg Met Ser Lys Asn Glu Phe Gly 195 200 205 Val Trp Glu Ile Phe Leu Pro Asn Asn Ala Asp Gly Thr Ser Pro Ile 210 215 220 Pro His Gly Ser Arg Val Lys Val Arg Met Asp Thr Pro Ser Gly Ile 225 230 235 240 Lys Asp Ser Ile Pro Ala Trp Ile Lys Tyr Ser Val Gln Ala Pro Gly 245 250 255 Glu Ile Pro Tyr Asp Gly Ile Tyr Tyr Asp Pro Pro Glu Glu Val Lys 260 265 270 Tyr Val Phe Arg His Ala Gln Pro Lys Arg Pro Lys Ser Leu Arg Ile 275 280 285 Tyr Glu Thr His Val Gly Met Ser Ser Pro Glu Pro Lys Ile Asn Thr 290 295 300 Tyr Val Asn Phe Arg Asp Glu Val Leu Pro Arg Ile Lys Lys Leu Gly 305 310 315 320 Tyr Asn Ala Val Gln Ile Met Ala Ile Gln Glu His Ser Tyr Tyr Gly 325 330 335 Ser Phe Gly Tyr His Val Thr Asn Phe Phe Ala Pro Ser Ser Arg Phe 340 345 350 Gly Thr Pro Glu Asp Leu Lys Ser Leu Ile Asp Arg Ala His Glu Leu 355 360 365 Gly Leu Leu Val Leu Met Asp Val Val His Ser His Ala Ser Ser Asn 370 375 380 Thr Leu Asp Gly Leu Asn Gly Phe Asp Gly Thr Asp Thr His Tyr Phe 385 390 395 400 His Ser Gly Pro Arg Gly His His Trp Met Trp Asp Ser Arg Leu Phe 405 410 415 Asn Tyr Gly Asn Trp Glu Val Leu Arg Phe Leu Leu Ser Asn Ala Arg 420 425 430 Trp Trp Leu Glu Glu Tyr Lys Phe Asp Gly Phe Arg Phe Asp Gly Val 435 440 445 Thr Ser Met Met Tyr Thr His His Gly Leu Gln Val Thr Phe Thr Gly 450 455 460 Asn Phe Asn Glu Tyr Phe Gly Phe Ala Thr Asp Val Asp Ala Val Val 465 470 475 480 Tyr Leu Met Leu Val Asn Asp Leu Ile His Gly Leu Tyr Pro Glu Ala 485 490 495 Val Thr Ile Gly Glu Asp Val Ser Gly Met Pro Thr Phe Ala Leu Pro 500 505 510 Val His Asp Gly Gly Val Gly Phe Asp Tyr Arg Met His Met Ala Val 515 520 525 Ala Asp Lys Trp Ile Asp Leu Leu Lys Gln Ser Asp Glu Thr Trp Lys 530 535 540 Met Gly Asp Ile Val His Thr Leu Thr Asn Arg Arg Trp Leu Glu Lys 545 550 555 560 Cys Val Thr Tyr Ala Glu Ser His Asp Gln Ala Leu Val Gly Asp Lys 565 570 575 Thr Ile Ala Phe Trp Leu Met Asp Lys Asp Met Tyr Asp Phe Met Ala 580 585 590 Leu Asp Arg Pro Ser Thr Pro Thr Ile Asp Arg Gly Ile Ala Leu His 595 600 605 Lys Met Ile Arg Leu Ile Thr Met Gly Leu Gly Gly Glu Gly Tyr Leu 610 615 620 Asn Phe Met Gly Asn Glu Phe Gly His Pro Glu Trp Ile Asp Phe Pro 625 630 635 640 Arg Gly Pro Gln Arg Leu Pro Ser Gly Lys Phe Ile Pro Gly Asn Asn 645 650 655 Asn Ser Tyr Asp Lys Cys Arg Arg Arg Phe Asp Leu Gly Asp Ala Asp 660 665 670 Tyr Leu Arg Tyr His Gly Met Gln Glu Phe Asp Gln Ala Met Gln His 675 680 685 Leu Glu Gln Lys Tyr Glu Phe Met Thr Ser Asp His Gln Tyr Ile Ser 690 695 700 Arg Lys His Glu Glu Asp Lys Val Ile Val Phe Glu Lys Gly Asp Leu 705 710 715 720 Val Phe Val Phe Asn Phe His Cys Asn Asn Ser Tyr Phe Asp Tyr Arg 725 730 735 Ile Gly Cys Arg Lys Pro Gly Val Tyr Lys Val Val Leu Asp Ser Asp 740 745 750 Ala Gly Leu Phe Gly Gly Phe Ser Arg Ile His His Ala Ala Glu His 755 760 765 Phe Thr Ala Asp Cys Ser His Asp Asn Arg Pro Tyr Ser Phe Ser Val 770 775 780 Tyr Thr Pro Ser Arg Thr Cys Val Val Tyr Ala Pro Val Glu Arg Gly 785 790 795 800 Thr Arg Cys Cys Ala Ala Cys Val Gly Leu Ser Met Gly Lys Thr Phe 805 810 815 Phe Gln Asn Arg Gln Met His Ala Cys Met Leu Gln Gly Ser Asp Thr 820 825 830 Leu Ile Asp Ala Gly Lys Pro Met His Leu Ala Ala Leu Ser Ser Leu 835 840 845 Phe Ile Asp Leu Gln Gly Val Asn Thr Ser Phe Arg Phe Ser Leu Lys 850 855 860 Lys Lys Lys Lys Lys Leu 865 870 55 2640 DNA Zea mays 55 gcgagatggc gttccgggtt tctggggcgg tgctcggtgg ggccgtaagg gctccccgac 60 tcaccggcgg cggggagggt agtctagtct tccggcacac cggcctcttc ttaactcggg 120 gtgctcgagt tggatgttcg gggacgcacg gggccatgcg cgcggcggcc gcggccagga 180 aggcggtcat ggttcctgag ggcgagaatg atggcctcgc atcaagggct gactcggctc 240 aattccagtc ggatgaactg gaggtaccag acatttctga agagacaacg tgcggtgctg 300 gtgtggctga tgctcaagcc ttgaacagag ttcgagtggt ccccccacca agcgatggac 360 aaaaaatatt ccagattgac cccatgttgc aaggctataa gtaccatctt gagtatcggt 420 acagcctcta tagaagaatc cgttcagaca ttgatgaaca tgaaggaggc ttggaagcct 480 tctcccgtag ttatgagaag tttggattta atgccagcgc ggaaggtatc acatatcgag 540 aatgggctcc tggagcattt tctgcagcat tggtgggtga cgtcaacaac tgggatccaa 600 atgcagatcg tatgagcaaa aatgagtttg gtgtttggga aatttttctg cctaacaatg 660 cagatggtac atcacctatt cctcatggat ctcgtgtaaa ggtgagaatg gatactccat 720 cagggataaa ggattcaatt ccagcctgga tcaagtactc agtgcaggcc ccaggagaaa 780 taccatatga tgggatttat tatgatcctc ctgaagaggt aaagtatgtg ttcaggcatg 840 cgcaacctaa acgaccaaaa tcattgcgga tatatgaaac acatgtcgga atgagtagcc 900 cggaaccgaa gataaacaca tatgtaaact ttagggatga agtcctccca agaataaaaa 960 aacttggata caatgcagtg caaataatgg caatccaaga gcactcatat tatggaagct 1020 ttggatacca tgtaactaat ttttttgcgc caagtagtcg ttttggtacc ccagaagatt 1080 tgaagtcttt gattgataga gcacatgagc ttggtttgct agttctcatg gatgtggttc 1140 atagtcatgc gtcaagtaat actctggatg ggttgaatgg ttttgatggt acagatacac 1200 attactttca cagtggtcca cgtggccatc actggatgtg ggattctcgc ctatttaact 1260 atgggaactg ggaagtttta agatttcttc tctccaatgc tagatggtgg ctcgaggaat 1320 ataagtttga tggtttccgt tttgatggtg tgacctccat gatgtacact caccacggat 1380 tacaagtaac atttacgggg aacttcaatg agtattttgg ctttgccacc gatgtagatg 1440 cagtggttta cttgatgctg gtaaatgatc taattcatgg actttatcct gaggctgtaa 1500 ccattggtga agatgttagt ggaatgccta catttgccct tcctgttcac gatggtgggg 1560 taggttttga ctatcggatg catatggctg tggctgacaa atggattgac cttctcaagc 1620 aaagtgatga aacttggaag atgggtgata ttgtgcacac actgacaaat aggaggtggt 1680 tagagaagtg tgtaacttat gctgaaagtc atgatcaagc attagtcggc gacaagacta 1740 ttgcgttttg gttgatggac aaggatatgt atgatttcat ggccctcgat agaccttcaa 1800 ctcctaccat tgatcgtggg atagcattac ataagatgat tagacttatc acaatgggtt 1860 taggaggaga gggctatctt aatttcatgg gaaatgagtt tggacatcct gaatggatag 1920 attttccaag aggtccgcaa agacttccaa gtggtaagtt tattccaggg aataacaaca 1980 gttatgacaa atgtcgtcga agatttgacc tgggtgatgc agactatctt aggtatcatg 2040 gtatgcaaga gtttgatcag gcaatgcaac atcttgagca aaaatatgaa ttcatgacat 2100 ctgatcacca gtatatttcc cggaaacatg aggaggataa ggtgattgtg ttcgaaaagg 2160 gagatttggt atttgtgttc aacttccact gcaacaacag ctattttgac taccgtattg 2220 gttgtcgaaa gcctggggtg tataaggtgg tcttggactc cgacgctgga ctatttggtg 2280 gatttagcag gatccatcac gcagccgagc acttcaccgc cgactgttcg catgataata 2340 ggccatattc attctcggtt tatacaccaa gcagaacatg tgtcgtctat gctccagtgg 2400 agtgatagcg gggtactcgt tgctgcgcgg catgtgtggg gctgtcgatg tgaggaaaaa 2460 ccttcttcca aaaccggcag atgcatgcat gcatgctaca ataaggttct gatactttaa 2520 tcgatgctgg aaagcccatg catctcgctg cgttgtcctc tctatttata taagaccttc 2580 aaggtgtcaa ttaaacatag agttttcgtt tttcgcttaa aaaaaaaaaa aaaaaactca 2640 56 776 PRT Zea mays 56 Ala Thr Val Gln Glu Asp Lys Thr Met Ala Thr Ala Lys Gly Asp Val 1 5 10 15 Asp His Leu Pro Ile Tyr Asp Leu Asp Pro Lys Leu Glu Ile Phe Lys 20 25 30 Asp His Phe Arg Tyr Arg Met Lys Arg Phe Leu Glu Gln Lys Gly Ser 35 40 45 Ile Glu Glu Asn Glu Gly Ser Leu Glu Ser Phe Ser Lys Gly Tyr Leu 50 55 60 Lys Phe Gly Ile Asn Thr Asn Glu Asp Gly Thr Val Tyr Arg Glu Trp 65 70 75 80 Ala Pro Ala Ala Gln Glu Ala Glu Leu Ile Gly Asp Phe Asn Asp Trp 85 90 95 Asn Gly Ala Asn His Lys Met Glu Lys Asp Lys Phe Gly Val Trp Ser 100 105 110 Ile Lys Ile Asp His Val Lys Gly Lys Pro Ala Ile Pro His Asn Ser 115 120 125 Lys Val Lys Phe Arg Phe Leu His Gly Gly Val Trp Val Asp Arg Ile 130 135 140 Pro Ala Leu Ile Arg Tyr Ala Thr Val Asp Ala Ser Lys Phe Gly Ala 145 150 155 160 Pro Tyr Asp Gly Val His Trp Asp Pro Pro Ala Ser Glu Arg Tyr Thr 165 170 175 Phe Lys His Pro Arg Pro Ser Lys Pro Ala Ala Pro Arg Ile Tyr Glu 180 185 190 Ala His Val Gly Met Ser Gly Glu Lys Pro Ala Val Ser Thr Tyr Arg 195 200 205 Glu Phe Ala Asp Asn Val Leu Pro Arg Ile Arg Ala Asn Asn Tyr Asn 210 215 220 Thr Val Gln Leu Met Ala Val Met Glu His Ser Tyr Tyr Ala Ser Phe 225 230 235 240 Gly Tyr His Val Thr Asn Phe Phe Ala Val Ser Ser Arg Ser Gly Thr 245 250 255 Pro Glu Asp Leu Lys Tyr Leu Val Asp Lys Ala His Ser Leu Gly Leu 260 265 270 Arg Val Leu Met Asp Val Val His Ser His Ala Ser Asn Asn Val Thr 275 280 285 Asp Gly Leu Asn Gly Tyr Asp Val Gly Gln Ser Thr Gln Glu Ser Tyr 290 295 300 Phe His Ala Gly Asp Arg Gly Tyr His Lys Leu Trp Asp Ser Arg Leu 305 310 315 320 Phe Asn Tyr Ala Asn Trp Glu Val Leu Arg Phe Leu Leu Ser Asn Leu 325 330 335 Arg Tyr Trp Leu Asp Glu Phe Met Phe Asp Gly Phe Arg Phe Asp Gly 340 345 350 Val Thr Ser Met Leu Tyr His His His Gly Ile Asn Val Gly Phe Thr 355 360 365 Gly Asn Tyr Gln Glu Tyr Phe Ser Leu Asp Thr Ala Val Asp Ala Val 370 375 380 Val Tyr Met Met Leu Ala Asn His Leu Met His Lys Leu Leu Pro Glu 385 390 395 400 Ala Thr Val Val Ala Glu Asp Val Ser Gly Met Pro Val Leu Cys Arg 405 410 415 Pro Val Asp Glu Gly Gly Val Gly Phe Asp Tyr Arg Leu Ala Met Ala 420 425 430 Ile Pro Asp Arg Trp Ile Asp Tyr Leu Lys Asn Lys Asp Asp Ser Glu 435 440 445 Trp Ser Met Gly Glu Ile Ala His Thr Leu Thr Asn Arg Arg Tyr Thr 450 455 460 Glu Lys Cys Ile Ala Tyr Ala Glu Ser His Asp Gln Ser Ile Val Gly 465 470 475 480 Asp Lys Thr Ile Ala Phe Leu Leu Met Asp Lys Glu Met Tyr Thr Gly 485 490 495 Met Ser Asp Leu Gln Pro Ala Ser Pro Thr Ile Asp Arg Gly Ile Ala 500 505 510 Leu Gln Lys Met Ile His Phe Ile Thr Met Ala Leu Gly Gly Asp Gly 515 520 525 Tyr Leu Asn Phe Met Gly Asn Glu Phe Gly His Pro Glu Trp Ile Asp 530 535 540 Phe Pro Arg Glu Gly Asn Asn Trp Ser Tyr Asp Lys Cys Arg Arg Gln 545 550 555 560 Trp Ser Leu Val Asp Thr Asp His Leu Arg Tyr Lys Tyr Met Asn Ala 565 570 575 Phe Asp Gln Ala Met Asn Ala Leu Asp Glu Arg Phe Ser Phe Leu Ser 580 585 590 Ser Ser Lys Gln Ile Val Ser Asp Met Asn Asp Glu Glu Lys Val Ile 595 600 605 Val Phe Glu Arg Gly Asp Leu Val Phe Val Phe Asn Phe His Pro Lys 610 615 620 Lys Thr Tyr Glu Gly Tyr Lys Val Gly Cys Asp Leu Pro Gly Lys Tyr 625 630 635 640 Arg Val Ala Leu Asp Ser Asp Ala Leu Val Phe Gly Gly His Gly Arg 645 650 655 Val Gly His Asp Val Asp His Phe Thr Ser Pro Glu Gly Val Pro Gly 660 665 670 Val Pro Glu Thr Asn Phe Asn Asn Arg Pro Asn Ser Phe Lys Val Leu 675 680 685 Ser Pro Pro Arg Thr Cys Val Ala Tyr Tyr Arg Val Asp Glu Ala Gly 690 695 700 Ala Gly Arg Arg Leu His Ala Lys Ala Glu Thr Gly Lys Thr Ser Pro 705 710 715 720 Ala Glu Ser Ile Asp Val Lys Ala Ser Arg Ala Ser Ser Lys Glu Asp 725 730 735 Lys Glu Ala Thr Ala Gly Gly Lys Lys Gly Trp Lys Phe Ala Arg Gln 740 745 750 Pro Ser Asp Gln Asp Thr Lys Trp Cys Leu Ile Thr Glu Gln Ala Gly 755 760 765 Thr Ala Cys Ile Ala Phe Leu Glu 770 775 57 2763 DNA Zea mays 57 gctgtgcctc gtgtcgccct cttcctcgcc gactccgctt ccgccgccgc ggcgctctcg 60 ctcgcatgct gatcgggcgg caccgccggg gatcgcgggt ggcggcaatg tgcgcctgag 120 tgtgttgtct gtccagtgca aggctcgccg gtcaggggtg cggaaggtca agagcaaatt 180 cgccactgca gctactgtgc aagaagataa aactatggca actgccaaag gcgatgtcga 240 ccatctcccc atatacgacc tggaccccaa gctggagata ttcaaggacc atttcaggta 300 ccggatgaaa agattcctag agcagaaagg atcaattgaa gaaaatgagg gaagtcttga 360 atctttttct aaaggctatt tgaaatttgg gattaataca aatgaggatg gaactgtata 420 tcgtgaatgg gcacctgctg cgcaggaggc agagcttatt ggtgacttca atgactggaa 480 tggtgcaaac cataagatgg agaaggataa atttggtgtt tggtcgatca aaattgacca 540 tgtcaaaggg aaacctgcca tccctcacaa ttccaaggtt aaatttcgct ttctacatgg 600 tggagtatgg gttgatcgta ttccagcatt gattcgttat gcgactgttg atgcctctaa 660 atttggagct ccctatgatg gtgttcattg ggatcctcct gcttctgaaa ggtacacatt 720 taagcatcct cggccttcaa agcctgctgc tccacgtatc tatgaagccc atgtaggtat 780 gagtggtgaa aagccagcag taagcacata tagggaattt gcagacaatg tgttgccacg 840 catacgagca aataactaca acacagttca gttgatggca gttatggagc attcgtacta 900 tgcttctttc gggtaccatg tgacaaattt ctttgcggtt agcagcagat caggcacacc 960 agaggacctc aaatatcttg ttgataaggc acacagtttg ggtttgcgag ttctgatgga 1020 tgttgtccat agccatgcaa gtaataatgt cacagatggt ttaaatggct atgatgttgg 1080 acaaagcacc caagagtcct attttcatgc gggagataga ggttatcata aactttggga 1140 tagtcggctg ttcaactatg ctaactggga ggtattaagg tttcttcttt ctaacctgag 1200 atattggttg gatgaattca tgtttgatgg cttccgattt gatggagtta catcaatgct 1260 gtatcatcac catggtatca atgtggggtt tactggaaac taccaggaat atttcagttt 1320 ggacacagct gtggatgcag ttgtttacat gatgcttgca aaccatttaa tgcacaaact 1380 cttgccagaa gcaactgttg ttgctgaaga tgtttcaggc atgccggtcc tttgccggcc 1440 agttgatgaa ggtggggttg ggtttgacta tcgcctggca atggctatcc ctgatagatg 1500 gattgactac ctgaagaata aagatgactc tgagtggtcg atgggtgaaa tagcgcatac 1560 tttgactaac aggagatata ctgaaaaatg catcgcatat gctgagagcc atgatcagtc 1620 tattgttggc gacaaaacta ttgcatttct cctgatggac aaggaaatgt acactggcat 1680 gtcagacttg cagcctgctt cacctacaat tgatcgaggg attgcactcc aaaagatgat 1740 tcacttcatc acaatggccc ttggaggtga tggctacttg aattttatgg gaaatgagtt 1800 tggtcaccca gaatggattg actttccaag agaagggaac aactggagct atgataaatg 1860 cagacgacag tggagccttg tggacactga tcacttgcgg tacaagtaca tgaatgcgtt 1920 tgaccaagcg atgaatgcgc tcgatgagag attttccttc ctttcgtcgt caaagcagat 1980 cgtcagcgac atgaacgatg aggaaaaggt tattgtcttt gaacgtggag atttagtttt 2040 tgttttcaat ttccatccca agaaaactta cgagggctac aaagtgggat gcgatttgcc 2100 tgggaaatac agagtagccc tggactctga tgctctggtc ttcggtggac atggaagagt 2160 tggccacgac gtggatcact tcacgtcgcc tgaaggggtg ccaggggtgc ccgaaacgaa 2220 cttcaacaac cggccgaact cgttcaaagt cctttctccg ccccgcacct gtgtggctta 2280 ttaccgtgta gacgaagcag gggctggacg acgtcttcac gcgaaagcag agacaggaaa 2340 gacgtctcca gcagagagca tcgacgtcaa agcttccaga gctagtagca aagaagacaa 2400 ggaggcaacg gctggtggca agaagggatg gaagtttgcg cggcagccat ccgatcaaga 2460 taccaaatga agccacgagt ccttggtgag gactggactg gctgccggcg ccctgttagt 2520 agtcctgctc tactggacta gccgccgctg gcgcccttgg aacggtcctt tcctgtagct 2580 tgcaggcgac tggtgtctca tcaccgagca ggcaggcact gcttgtatag cttttctaga 2640 ataataatca gggatggatg gatggtgtgt attggctatc tggctagacg tgcatgtgcc 2700 cagtttgtat gtacaggagc agttcccgtc cagaataaaa aaaaacttgt tggggggttt 2760 ttc 2763 58 153 DNA Zea mays CDS (1)..(153) 58 atg gcg acg ccc tcg gcc gtg ggc gcc gcg tgc ctc ctc ctc gcg cgg 48 Met Ala Thr Pro Ser Ala Val Gly Ala Ala Cys Leu Leu Leu Ala Arg 1 5 10 15 gcc gcc tgg ccg gcc gcc gtc ggc gac cgg gcg cgc ccg cgg cgg ctc 96 Ala Ala Trp Pro Ala Ala Val Gly Asp Arg Ala Arg Pro Arg Arg Leu 20 25 30 cag cgc gtg ctg cgc cgc cgg tgc gtc gcg gag ctg agc agg gag ggg 144 Gln Arg Val Leu Arg Arg Arg Cys Val Ala Glu Leu Ser Arg Glu Gly 35 40 45 ccc cat atg 153 Pro His Met 50 59 51 PRT Zea mays 59 Met Ala Thr Pro Ser Ala Val Gly Ala Ala Cys Leu Leu Leu Ala Arg 1 5 10 15 Ala Ala Trp Pro Ala Ala Val Gly Asp Arg Ala Arg Pro Arg Arg Leu 20 25 30 Gln Arg Val Leu Arg Arg Arg Cys Val Ala Glu Leu Ser Arg Glu Gly 35 40 45 Pro His Met 50 60 9 PRT Zea mays MOD_RES (7) Arg or Asp 60 Cys Val Ala Glu Leu Ser Xaa Leu Gly 1 5 61 8 PRT Zea mays 61 Gly Glu Asn Val Met Asn Val Ile 1 5 62 10 PRT Zea mays 62 Cys Val Ala Glu Leu Ser Arg Glu Gly Pro 1 5 10 63 8 PRT Zea mays 63 Ser Ile Val Phe Val Thr Gly Glu 1 5 64 8 PRT Zea mays 64 Ala Ser Ala Gly Met Asn Val Val 1 5 65 8 PRT Zea mays 65 Ala Glu Ala Glu Ala Gly Gly Lys 1 5 66 8 PRT Zea mays 66 Gly Glu Asn Val Met Asn Val Ile 1 5 67 9 PRT Zea mays 67 Gly Ser Val Gly Ala Ala Leu Arg Ser 1 5 68 8 PRT Zea mays 68 Ala Ala Ala Pro Ala Gly Glu Glu 1 5 69 8 PRT Zea mays 69 Met Val Val Val Trp Ala Ser Glu 1 5 70 798 PRT Zea mays 70 Met Ala Phe Arg Val Ser Gly Ala Val Leu Gly Gly Ala Val Arg Ala 1 5 10 15 Pro Arg Leu Thr Gly Gly Gly Glu Gly Ser Leu Val Phe Arg His Thr 20 25 30 Gly Leu Phe Leu Thr Arg Gly Ala Arg Val Gly Cys Ser Gly Thr His 35 40 45 Gly Ala Met Arg Ala Ala Ala Ala Ala Arg Lys Ala Met Val Pro Glu 50 55 60 Gly Glu Asn Asp Gly Leu Ala Ser Arg Ala Asp Ser Ala Gln Phe Gln 65 70 75 80 Ser Asp Glu Leu Glu Val Pro Asp Ile Ser Glu Glu Thr Thr Cys Gly 85 90 95 Ala Gly Val Ala Asp Ala Gln Ala Leu Asn Arg Val Arg Val Val Pro 100 105 110 Pro Pro Ser Asp Gly Gln Lys Ile Phe Gln Ile Asp Pro Met Leu Gln 115 120 125 Gly Tyr Lys Tyr His Leu Glu Tyr Arg Tyr Ser Leu Tyr Arg Arg Ile 130 135 140 Arg Ser Asp Ile Asp Glu His Glu Gly Gly Leu Glu Ala Phe Ser Arg 145 150 155 160 Ser Tyr Glu Lys Phe Gly Phe Asn Ala Ser Ala Glu Gly Ile Thr Tyr 165 170 175 Arg Glu Trp Ala Pro Gly Ala Phe Ser Ala Ala Leu Val Gly Asp Val 180 185 190 Asn Asn Trp Asp Pro Asn Ala Asp Arg Met Ser Lys Asn Glu Phe Gly 195 200 205 Val Trp Glu Ile Phe Leu Pro Asn Asn Ala Asp Gly Thr Ser Pro Ile 210 215 220 Pro His Gly Ser Arg Val Lys Val Arg Met Asp Thr Pro Ser Gly Ile 225 230 235 240 Lys Asp Ser Ile Pro Ala Trp Ile Lys Tyr Ser Val Gln Ala Pro Gly 245 250 255 Glu Ile Pro Tyr Asp Gly Ile Tyr Tyr Asp Pro Pro Glu Glu Val Lys 260 265 270 Tyr Val Phe Arg His Ala Gln Pro Lys Arg Pro Lys Ser Leu Arg Ile 275 280 285 Tyr Glu Thr His Val Gly Met Ser Ser Pro Glu Pro Lys Ile Asn Thr 290 295 300 Tyr Val Asn Phe Arg Asp Glu Val Leu Pro Arg Ile Lys Lys Leu Gly 305 310 315 320 Tyr Asn Ala Val Gln Ile Met Ala Ile Gln Glu His Ser Tyr Tyr Gly 325 330 335 Ser Phe Gly Tyr His Val Thr Asn Phe Phe Ala Pro Ser Ser Arg Phe 340 345 350 Gly Thr Pro Glu Asp Leu Lys Ser Leu Ile Asp Arg Ala His Glu Leu 355 360 365 Gly Leu Leu Val Leu Met Asp Val Val His Ser His Ala Ser Ser Asn 370 375 380 Thr Leu Asp Gly Leu Asn Gly Phe Asp Gly Thr Asp Thr His Tyr Phe 385 390 395 400 His Ser Gly Pro Arg Gly His His Trp Met Trp Asp Ser Arg Leu Phe 405 410 415 Asn Tyr Gly Asn Trp Glu Val Leu Arg Phe Leu Leu Ser Asn Ala Arg 420 425 430 Trp Trp Leu Glu Glu Tyr Lys Phe Asp Gly Phe Arg Phe Asp Gly Val 435 440 445 Thr Ser Met Met Tyr Thr His His Gly Leu Gln Val Thr Phe Thr Gly 450 455 460 Asn Phe Asn Glu Tyr Phe Gly Phe Ala Thr Asp Val Asp Ala Val Val 465 470 475 480 Tyr Leu Met Leu Val Asn Asp Leu Ile His Gly Leu Tyr Pro Glu Ala 485 490 495 Val Thr Ile Gly Glu Asp Val Ser Gly Met Pro Thr Phe Ala Leu Pro 500 505 510 Val His Asp Gly Gly Val Gly Phe Asp Tyr Arg Met His Met Ala Val 515 520 525 Ala Asp Lys Trp Ile Asp Leu Leu Lys Gln Ser Asp Glu Thr Trp Lys 530 535 540 Met Gly Asp Ile Val His Thr Leu Thr Asn Arg Arg Trp Leu Glu Lys 545 550 555 560 Cys Val Thr Tyr Ala Glu Ser His Asp Gln Ala Leu Val Gly Asp Lys 565 570 575 Thr Ile Ala Phe Trp Leu Met Asp Lys Asp Met Tyr Asp Phe Met Ala 580 585 590 Leu Asp Arg Pro Ser Thr Pro Thr Ile Asp Arg Gly Ile Ala Leu His 595 600 605 Lys Met Ile Arg Leu Ile Thr Met Gly Leu Gly Gly Glu Gly Tyr Leu 610 615 620 Asn Phe Met Gly Asn Glu Phe Gly His Pro Glu Trp Ile Asp Phe Pro 625 630 635 640 Arg Gly Pro Gln Arg Leu Pro Ser Gly Lys Phe Ile Pro Gly Asn Asn 645 650 655 Asn Ser Tyr Asp Lys Cys Arg Arg Arg Phe Asp Leu Gly Asp Ala Asp 660 665 670 Tyr Leu Arg Tyr His Gly Met Gln Glu Phe Asp Gln Ala Met Gln His 675 680 685 Leu Glu Gln Lys Tyr Glu Phe Met Thr Ser Asp His Gln Tyr Ile Ser 690 695 700 Arg Lys His Glu Glu Asp Lys Val Ile Val Phe Glu Lys Gly Asp Leu 705 710 715 720 Val Phe Val Phe Asn Phe His Cys Asn Asn Ser Tyr Phe Asp Tyr Arg 725 730 735 Ile Gly Cys Arg Lys Pro Gly Val Tyr Lys Val Val Leu Asp Ser Asp 740 745 750 Ala Gly Leu Phe Gly Gly Phe Ser Arg Ile His His Ala Ala Glu His 755 760 765 Phe Thr Ala Asp Cys Ser His Asp Asn Arg Pro Tyr Ser Phe Ser Val 770 775 780 Tyr Thr Pro Ser Arg Thr Cys Val Val Tyr Ala Pro Val Glu 785 790 795 71 14 PRT Zea mays 71 Arg Gly Thr Arg Cys Cys Ala Ala Cys Val Gly Leu Ser Met 1 5 10 72 16 PRT Zea mays 72 Gly Lys Thr Phe Phe Gln Asn Arg Gln Met His Ala Cys Met Leu Gln 1 5 10 15 73 22 PRT Zea mays 73 Gly Ser Asp Thr Leu Ile Asp Ala Gly Lys Pro Met His Leu Ala Ala 1 5 10 15 Leu Ser Ser Leu Phe Ile 20 74 6 PRT Zea mays 74 Asp Leu Gln Gly Val Asn 1 5 75 13 PRT Zea mays 75 Ser Phe Arg Phe Ser Leu Lys Lys Lys Lys Lys Lys Leu 1 5 10 76 431 PRT E. coli glgC3 76 Met Val Ser Leu Glu Lys Asn Asp His Leu Met Leu Ala Arg Gln Leu 1 5 10 15 Pro Leu Lys Ser Val Ala Leu Ile Leu Ala Gly Gly Arg Gly Thr Arg 20 25 30 Leu Lys Asp Leu Thr Asn Lys Arg Ala Lys Pro Ala Val His Phe Gly 35 40 45 Gly Lys Phe Arg Ile Ile Asp Phe Ala Leu Ser Asn Cys Ile Asn Ser 50 55 60 Gly Ile Arg Arg Met Gly Val Ile Thr Gln Tyr Gln Ser His Thr Leu 65 70 75 80 Val Gln His Ile Gln Arg Gly Trp Ser Phe Phe Asn Glu Glu Met Asn 85 90 95 Glu Phe Val Asp Leu Leu Pro Ala Gln Gln Arg Met Lys Gly Glu Asn 100 105 110 Trp Tyr Arg Gly Thr Ala Asp Ala Val Thr Gln Asn Leu Asp Ile Ile 115 120 125 Arg Arg Tyr Lys Ala Glu Tyr Val Val Ile Leu Ala Gly Asp His Ile 130 135 140 Tyr Lys Gln Asp Tyr Ser Arg Met Leu Ile Asp His Val Glu Lys Gly 145 150 155 160 Val Arg Cys Thr Val Val Cys Met Pro Val Pro Ile Glu Glu Ala Ser 165 170 175 Ala Phe Gly Val Met Ala Val Asp Glu Asn Asp Lys Thr Ile Glu Phe 180 185 190 Val Glu Lys Pro Ala Asn Pro Pro Ser Met Pro Asn Asp Pro Ser Lys 195 200 205 Ser Leu Ala Ser Met Gly Ile Tyr Val Phe Asp Ala Asp Tyr Leu Tyr 210 215 220 Glu Leu Leu Glu Glu Asp Asp Arg Asp Glu Asn Ser Ser His Asp Phe 225 230 235 240 Gly Lys Asp Leu Ile Pro Lys Ile Thr Glu Ala Gly Leu Ala Tyr Ala 245 250 255 His Pro Phe Pro Leu Ser Cys Val Gln Ser Asp Pro Asp Ala Glu Pro 260 265 270 Tyr Trp Arg Asp Val Gly Thr Leu Glu Ala Tyr Trp Lys Ala Asn Leu 275 280 285 Asp Leu Ala Ser Val Val Asp Lys Leu Asp Met Tyr Asp Arg Asn Trp 290 295 300 Pro Ile Arg Thr Tyr Asn Glu Ser Leu Pro Pro Ala Lys Phe Val Gln 305 310 315 320 Asp Arg Ser Gly Ser His Gly Met Thr Leu Asn Ser Leu Val Ser Gly 325 330 335 Gly Cys Val Ile Ser Gly Ser Val Val Val Gln Ser Val Leu Phe Ser 340 345 350 Arg Val Arg Val Asn Ser Phe Cys Asn Ile Asp Ser Ala Val Leu Leu 355 360 365 Pro Glu Val Trp Val Gly Arg Ser Cys Arg Leu Arg Arg Cys Val Ile 370 375 380 Asp Arg Ala Cys Val Ile Pro Glu Gly Met Val Ile Gly Glu Asn Ala 385 390 395 400 Glu Glu Asp Ala Arg Arg Phe Tyr Arg Ser Glu Glu Gly Ile Val Leu 405 410 415 Val Thr Arg Glu Met Leu Arg Lys Leu Gly His Lys Gln Glu Arg 420 425 430 77 10 PRT E. coli glgC3 77 Cys Arg Phe Tyr Met Tyr Val Gln Arg Cys 1 5 10 

What is claimed is:
 1. A method of producing polysaccharides which are non glycogen like in a host comprising: a transforming a host capable of being used in a fermentation process, with genes selected from the group which produce starch synthesizing enzymes, glycogen synthesizing enzymes such that the host produces nonglycogen like starch, and b. employing the host in a fermentation process wherein the fermentation process produces polysaccharides.
 2. A method according to claim 1 wherein the host is bacteria.
 3. A host transformed to carry a gene active in glycogen production, and at least one nonstarch branching gene active in the production of at least one of the following polysaccharides amylopectin and amylose in it original host.
 4. A host according to claim 3 wherein the host is a monocot.
 5. A host according to claim 3 wherein the host is a dicot.
 6. A host according to claim 3 wherein the host is a plant.
 7. A host according to claim 3 wherein the host is a bacteria.
 8. A host according to claim 3 wherein the host is a cereal bearing plant.
 9. A host according to claim 3 wherein the bacterial gene is selected from the group consisting of glgC gene, glgA gene, glgB gene.
 10. A host according to claim 3 wherein at least one nonstarch branching genes active in the production of at least one of the following polysaccharides amylopectin and amylose in it original plant is selected from the group consisting of starch soluble starch synthase I, II, III genes and debranching enzyme gene (sul), GBSS gene, sh2 gene and bt2 gene.
 11. A host according to claim 3 including at least one of the starch branching enzyme genes.
 12. A host according to claim 11 including the starch branching enzyme gene BEI gene.
 13. A host according to claim 11 including the starch branching enzyme gene BEII gene.
 14. A host according to claim 12 including the starch branching enzyme gene BEII gene.
 15. A host transformed to carry a gene active in ADPG production, and at least one starch gene active in the production of at least one of the following polysaccharides amylopectin and amylose in it original host wherein the host produces polysaccharides that are plant like starch and not glycogen like.
 16. A host transformed to carry a pyrophosphatase gene, glycogen synthase gene,
 17. A host according to claim 1 wherein the gene active in glycogen production is a bacterial gene.
 18. A host deficient in alpha 1,4 glucan synthesizing ability and alpha 1,4-1,6 branching enzyme capability transformed to express at least one a plant starch soluble synthesis gene.
 19. A host according to claim 18 including being transformed to express at least one gene encoding for debranching enzyme.
 20. A host according to claim 18 wherein said gene is encoding for starch soluble synthase enzyme
 1. 21. A host according to claim 18 wherein said gene is encoding for starch soluble synthase enzyme II.
 22. A host according to claim 18 wherein said gene is encoding for starch soluble synthase enzyme III.
 23. A host according to claim 18 including being transformed to express at least one gene encoding for starch branching enzyme.
 24. A host according to claim 23 wherein the starch branching enzyme is BEI.
 25. A host according to claim 23 wherein the starch branching enzyme is BEII.
 26. A plasmid wherein said plasmid is in a carrier host and said plasmid contains the SSII gene with the n terminus GENVMNVIVV wherein the gene is approximately 1561 base pairs in length.
 27. A Plasmid according to claim on e wherein said host is a bacterial host.
 28. A host according to claim two wherein the host is a wild type E. coli.
 29. A new polysaccharide produced by a transformed host comprising: said host having a wildtype, said wildtype of said host does not produce said new polysaccharide, said transformed host expressing at least two exogenous starch synthesis genes, said genes are selected from a group consisting of soluble starch synthesis genes such as SSI SSII SSIII, wherein the transformed host is capable of producing such new polysaccharide.
 30. The new polysaccharide of claim 29 wherein said host also expresses the exogenous genes selected from the following group consisting of bacterial glycogen inducing genes.
 31. The new polysaccharide of claim 29 wherein said host also expresses the exogenous genes selected from the following group consisting of plant granule bound enzymes.
 32. A method according to claim one wherein the host is fungal.
 33. A method according to claim 32 wherein the host is yeast.
 34. A method according to claim one wherein said glycogen synthesizing genes include glgC, glgA, glgB genes.
 35. A method according to claim one wherein said genes which produce starch synthesizing enzymes include genesencoding for starch soluble synthases I, II, III.
 36. A method according to claim one wherein said genes which produce starch synthesizing enzymes include genes encoding for and debranching enzyme and branching enzymes.
 37. The new polysaccharide of claim 30 wherein said bacterial glycogen inducing genes are selected from the group consisting of glgC, glgA, glgB.
 38. The new polysaccharide of claim 29 wherein said starch synthesis genes are selected from the group consisting of BEI and BEII. 